Pegylated alpha naphthol compositions and methods of treatment

Abstract

Disclosed, in part, are isolated, stable and biologically active pegylated menaphthol derivatives and methods of their use in the treatment of various diseases.

Description

[0001] Related Applications

[0002] This application is related to U.S. Provisional Patent Application No. 63 / 347,007 filed on May 30, 2022; U.S. Provisional Patent Application No. 63 / 347,008 filed on May 30, 2022; U.S. Provisional Patent Application No. 63 / 413,495 filed on October 5, 2022; U.S. Provisional Patent Application No. 63 / 413,506 filed on October 5, 2022; U.S. Provisional Patent Application No. 63 / 447,310 filed on February 21, 2023; and U.S. Provisional Patent Application No. 63 / 447,314 filed on February 21, 2023. Technical Field

[0003] The present invention relates to biologically active menaquinol derivatives, compositions and formulations and combinations thereof for treating diseases associated with vitamin K, its reduced and biologically active form menaquinol and its salts, including osteoporosis and osteopenia. Background Art

[0004] Vitamin K is a group of fat-soluble vitamins with similar structures. Vitamin K2, or menaquinone, has nine related compounds that can be subdivided into short-chain menaquinones (such as menaquinone-4 or MK-4) and long-chain menaquinones, such as MK-7, MK-8, and MK-9 to MK-12. The vitamins include phylloquinone (K1), menaquinone (K2), and menadione (K3). Plants synthesize vitamin K1, while bacteria can produce a range of vitamin K2 forms, including bacteria in the small intestine that convert K1 to K2. Vitamin K3 is a synthetic version of the vitamin and has been banned for human use by the US FDA due to its toxicity.

[0005] It has been shown that taking broad-spectrum antibiotics can reduce a person's intestinal production of vitamin K by nearly 74% compared to people who do not take these antibiotics. A diet low in vitamin K can also reduce the body's vitamin K concentration. Vitamin K1 is preferentially used by the liver as a clotting factor. Vitamin K2 is preferentially used by the brain, vascular system, breasts, and kidneys. Vitamin K2 aids in the production of myelin and sphingolipids (fats that are essential for brain health) and protects the brain from oxidative damage. Vitamin K2, as MK-4, promotes bone health by stimulating the production of connective tissue in the bones.

[0006] Vitamin K2 is the main storage form in animals, and there are several subtypes, which differ in that the chain length of the isoprenoid group or residue in the side chain is different. These vitamin K2 homologs are called menaquinones and are characterized by the number of isoprenoid residues in their side chains. For example, MK-4 has four isoprenoid residues in its side chain and is the most common type of vitamin K2 in animal products. MK-4 is usually synthesized from vitamin K1 in certain animal tissues (arterial walls, pancreas and testicles) by replacing the phytate group with an unsaturated geranyl group containing four isoprene units. Unlike MK-4, MK-7 is not produced by human tissues. MK-7 may be converted from phylloquinone (K1) in the colon by Escherichia coli (E.coli) bacteria. MK-4 and MK-7 are sold in the United States as bone health dietary supplements. MK-4 has been shown to reduce the incidence of fractures. MK-4, at a daily dose of 45 mg, has been approved by the Ministry of Health in Japan since 1995 for the prevention and treatment of osteoporosis.

[0007]

[0008] Vitamin K2, usually MK-4, has been proposed and evaluated in vitro and in vivo for the treatment of various cancers. For example, (1) Lamson et al., "The Anticancer Effects of Vitamin K", Altern. Med. Rev., 8(3), 303-318 (2003) reviewed vitamin K therapy. It is stated that "both in vitro and in vivo studies have shown that vitamin K2 also exhibits anticancer effects. Many cancer cell lines (including liver cancer, colon cancer, leukemia, lung cancer, gastric cancer, lymphocytic cancer, nasopharyngeal cancer, breast cancer and oral cancer epidermoid cells) were screened and their activity was observed. They also review proliferation inhibition in HOS TE85 human osteosarcoma and MC3T3-E1 mouse osteoblast cell lines, C6 rat and RBR17T and T98G human glioma cell lines, myeloblast (ML1) and promyelocytic (HL60) lung cancer cell lines, and state that vitamin K2 analogs, particularly MK-4, induce apoptosis in isolated leukemia cells (post-myelodysplastic syndrome (MDS) and acute myeloid leukemia). They provide two case reports; and they also mention pilot studies conducted in Japan with K2 (MK-4) for the treatment of MDS and post-MDS acute myeloid leukemia (post-MDS AML) and a trial in Japan in hepatocellular carcinoma, both of which showed promising results. (2) Yoshida et al., “Apoptosis induction of vitamin K2 in lung carcinoma cell lines: The possibility of vitamin K2 therapy for lung cancer,” Int. J. Oncol., 23, 627-632 (2003), stated that “vitamin K2 (menadione-4, VK2) has been reported to exhibit apoptosis and differentiation induction effects on leukemia cells” and that “the clinical benefit of using VK2 to treat patients with acute leukemia and myelodysplastic syndrome has been demonstrated.” MK-4 was shown to have in vitro activity against seven lung cancer cell lines: small cell lung cancer (LU-139, LU-I30); lung squamous cell carcinoma (LC-AI, LC-l / sq), lung adenocarcinoma (PC-14, CCL185) and large cell carcinoma (IA-LM); and it was also shown that a suboptimal dose of MK-4 combined with cisplatin had growth inhibitory activity against LU-139 cells in vitro.(3) Ohtsuka et al., “Vitamin K2 Inhibits the Growth and Invasiveness of Hepatocellular Carcinoma Cells via Protein Kinase A Activation”, Hepatology, 40, 243-251 (2004) reported that MK-4 inhibited hepatocellular carcinoma cells (HepG2 and PRF / PLC / 5) in vitro and in vivo (mouse xenografts with PRF / PLC / 5). They also mentioned an ongoing human clinical trial, the interim results of which were favorable for MK-4 treatment to inhibit portal vein invasion of liver cancer. (4) Tokita et al., “Vitamin K2-induced antitumor effects via cell-cycle arrest and apoptosis in gastric cancer cell lines”, Int. J. Molecular Med., 17, 235-243 (2006), showed that when four gastric cancer cells (KATO When cell lines III (signet ring cell carcinoma), MKN7 (well-differentiated tubular adenocarcinoma), MKN74 (moderately differentiated tubular adenocarcinoma, liver metastasis) and FU97 (poorly differentiated adenocarcinoma, AFP production) were exposed to MK-4 in vitro, cell growth was inhibited in a dose-dependent manner, and when the same cell lines were treated with up to 10 mg / L of 5-fluorouracil or docetaxel, cell growth in the group supplemented with MK-4 was inhibited in a dose-dependent manner compared with the control group. (5) Kawakita et al., "Growth inhibitory effects of vitamin K2 on colon cancer cell lines via different types of cell death including autophagy and apoptosis" apoptosis), International Journal of Molecular Medicine, 23, 709-716 (2009), showed that MK-4 showed a dose-dependent inhibitory effect in COLO201 and PMC01 colon cancer cell lines in vitro, but had no inhibitory effect in the DLD-1 cell line.(6) Li et al., “Induction of apoptosis in hepatocellular carcinoma Smmc-7721 cells by vitamin K2 is associated with p53 and independent of the intrinsic apoptotic pathway,” Mol. Cell Biochem., 342, 125-131 (2010), stated that “vitamin K2 (VK2) can exert cytostatic effects in various human cancer cells” and showed that vitamin K2 inhibited the cell proliferation of Smmc-7721 hepatocellular carcinoma cells in vitro in a dose-dependent manner. (7) Samykutty et al., “Vitamin K2, a Naturally Occurring Menaquinone, Exerts Therapeutic Effects on Both Hormone-Dependent and Hormone-Independent Prostate Cancer Cells,” Evid.-based Complement. Altern. Med., 287358 (2013), showed that vitamin K2 was able to inhibit the viability of both androgen-dependent and androgen-independent prostate cancer cells in vitro (LNCaP, DU145, and 22RV1 cells) and in vivo (mouse xenografts with LNCaP, DU145, and 22RV1 cells).(8) Kiely et al., “Real-time cell analysis of the inhibitory effect of vitamin K2 on adhesion and proliferation of breast cancer cells,” Nutr. Res., 35(8), 736-743 (2015), stated that “the antitumor properties of VK derivatives have been reported in both hepatocellular carcinoma and glioblastoma,” and showed that MK-4 was observed to have a dose-dependent growth inhibitory effect in vitro in the triple-negative breast cancer cell line MDA-MB-231 and the HER2+ breast cancer cell line MDA-MB-453. (9) Duan et al., “Vitamin K2 Induces Mitochondria-Related Apoptosis in Human Bladder Cancer Cells via ROS and JNKp38 MAPK Signal Pathways,” PLoS ONE, 11(8), e0161886 (2016) stated that “To date, a large number of studies have shown that vitamin K2 can exhibit anticancer activity in various cancer cell lines, including leukemia, lung cancer, ovarian cancer, prostate cancer, and hepatocellular carcinoma,” and in their study, MK-4 inhibited human bladder cancer cell lines (T24, J82, and EJ) in vitro and in vivo (mouse xenografts). (10) Mishima, “A non-canonical vitamin K cycle is a potent ferroptosis suppressor,” Nature, 608, 708-790 (2022), states that “ferroptosis is a non-apoptotic form of cell death characterized by iron-dependent lipid peroxidation and plays a key role in susceptibility to organ damage, degenerative diseases, and refractory cancers” and that “the fully reduced form of vitamin K—a group of naphthoquinones including menadione and phylloquinone—confers strong anti-ferroptosis functions” and serves to “protect cells from harmful lipid peroxidation and ferroptosis.”

[0009] Osteoporosis is a bone disease that causes an increased risk of fractures. In osteoporosis, bone mineral density (BMD) decreases, bone microstructure is disordered, and the number and type of non-collagenous proteins in the bones change. The World Health Organization defines osteoporosis (female) as a bone mineral density that is 2.5 standard deviations below peak bone mass (i.e., an average healthy woman of 30 years old). Osteoporosis is most common in women after menopause (called postmenopausal osteoporosis). Osteoporosis may also develop in men, and may occur in anyone with a specific hormone imbalance and other chronic diseases, or due to drugs, particularly glucocorticoids (when this disease is called steroid or glucocorticoid-induced osteoporosis), and due to nutritional deficiency states or other metabolic disorders, such as secondary consequences of hyponatremia or cancer, osteoporosis may occur. Osteopenia is a condition in which bone mineral density is lower than normal, and many doctors believe that this is a precursor to osteoporosis.

[0010] In most cases of osteoporosis, the underlying mechanism is an imbalance between bone resorption and bone formation. The three main mechanisms by which osteoporosis develops include insufficient peak bone mass (insufficient bone mass and strength during growth), excessive bone resorption, and insufficient new bone formation during remodeling. Hormonal factors largely determine the rate of bone resorption; a lack of estrogen (e.g., due to menopause) increases bone resorption and reduces new bone deposition that normally occurs in weight-bearing bones. In addition to estrogen, calcium metabolism plays an important role in bone turnover, and deficiencies in calcium and vitamin D lead to impaired bone deposition; in addition, the parathyroid gland responds to low calcium levels by secreting parathyroid hormone, which increases bone resorption to ensure that there is enough calcium in the blood. Drugs used to treat osteoporosis include calcium, vitamin D, vitamin K, bisphosphonates, calcitonin, teriparatide, strontium ranelate, hormone replacement, and selective estrogen receptor modulators.

[0011] It has been confirmed that cardiovascular disease (CVD) is the most common cause of death in patients with chronic kidney disease (CKD). When compared with the general population, when CKD patients are treated with hemodialysis, the cause of death caused by CVD is about 10-20 times. In addition, it has been shown that vascular calcification and related arteriosclerosis are very common in the incidence of CVD. Therefore, the disclosed method of treatment can be suitable for treating peripheral arterial disease. In addition, fractures (such as vertebral fractures and other types of fractures) in CKD patients who undergo dialysis treatment have a 3-fold higher risk.

[0012] It has been confirmed that there is a direct correlation between the level of vitamin K in the patient's blood and the incidence of vascular calcification, bone density and bone strength. Therefore, supplemental use of vitamin K, such as MK-7 and its fat-soluble hydroquinone (naphthol) derivatives disclosed herein, can provide significant clinical benefits for reducing vascular calcification caused in part by arteriosclerosis, and increase bone mineralization or bone mineral density, which will help treat or prevent CVD, and treat or prevent bone diseases in patients with CKD.

[0013] It was determined that nutritional doses of MK-7 and MK-9 were well absorbed by the human body and thus significantly increased the level of MK-7 in serum.

[0014]

[0015] In one embodiment, the present application discloses novel bio-effective naphthol derivatives and methods for using the same in treating various diseases, such as naphthol-7 derivatives and naphthol-9 derivatives, including salts thereof.

[0016] The above examples and limitations of the related art are intended to be illustrative and not exclusive. Other limitations of the related art will be apparent to those skilled in the art after reading the specification and studying the drawings as provided herein. Summary of the invention

[0017] The present inventors recognize that there is still a need to design novel compounds and formulations thereof that are effective for the indications disclosed above.The following embodiments, aspects, and variations thereof are exemplary and illustrative and are not intended to be limiting in scope.

[0018] The present inventors further expect that the novel PEGylated naphthol derivatives of the next paragraph will show the same vitamin K2-like (or MK-4-like) activity in diseases in which vitamin K2 and MK-4 show activity, and have similar utility in treating these diseases.

[0019] In one embodiment, the present application discloses an isolated, stable and biologically active naphthol derivative having the formula I , II or III :

[0020]

[0021] in:

[0022] m is an integer from 2 to 15; 1 an integer from 3 to 15 when present;

[0023] n is 7, 8, 9, or 10;

[0024] R is H, or is selected from the group consisting of: -C(O)C1-C6 alkyl, -C(O)(CH2) q -OC(O)C1-C6 alkyl and -Z-(C=Y)(CH2) s -(OCH2CH2)-OR 1 ;

[0025] q is 1, 2, or 3; R 1 is H or -CH3; R 2 is H or C1-C3 alkyl;

[0026] s is 1, 2 or 3; Y is O, NR 2 or S; Z is -C(O)CH2O-, -C(O)CH2NH- or absent; and

[0027] Z 1 It is -C(O)CH2O-, -C(O)CH2NH- or does not exist.

[0028] In-style I , II or III In one variation of each of the naphthol derivatives of 1 Each of m is independently 2 to 30. In another variation, m or m 1 Each of m and m is independently 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15. In another variation, m and m 1 Each of m and m is independently 6, 7, 8, 9, 10 or 11. In another variation, m and m 1 Each of m is independently 8, 9 or 10. In another variation, m or m 1 Each of is independently 12, 14 or 15. In another variation, the C1-C6 alkyl group is -CH3 or -CH2CH3. In another variation, the PEG group (i.e., a polyethylene glycol group, represented as -(OCH2CH2) m -or-(CH2OCH2) m -) has an average molecular weight of 200 Da (Daltons), 300 Da, 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1 kDa (kilodaltons), 1.5 kDa, 2 kDa, 3 kDa, 4 kDa or 5 kDa.

[0029] In one aspect, the naphthol derivative has the formula Ia , Ib or Ic :

[0030]

[0031] in:

[0032] m is an integer from 2 to 15; n is 7, 8, 9 or 10;

[0033] R is H, or is selected from the group consisting of: -C(O)C1-C6 alkyl, -C(O)(CH2) q -OC(O)C1-C6 alkyl and -Z-(C=Y)(CH2) s -(OCH2CH2)-OR 1 ;

[0034] q is 1, 2 or 3; s is 1, 2 or 3; R 1 is H or -CH3; R 2 is H or C1-C3 alkyl;

[0035] Y is O, NR 2 or S; and Z is -C(O)CH2O-, -C(O)CH2NH- or is absent.

[0036] In-style Ia In a variation of the naphthol derivative, R is CH3C(O)-, m is 7, R 1 is -CH3, and n is 7. Ia In another variation, R is CH3C(O)-, m is 7, R 1 is -CH3, and n is 9.

[0037] On the other hand, the naphthol derivative has the formula Ia.1 , Ib.1 or Ic.1 :

[0038]

[0039] wherein: m is an integer from 8 to 12; n is 7, 8, 9 or 10; and R 1 It is H or -CH3.

[0040] On the other hand, the naphthol derivative has the formula IIa , IIb or IIc :

[0041]

[0042] wherein: m is an integer from 8 to 12; n is 7, 8, 9 or 10; s is 1, 2 or 3; and R 1 is H or -CH3. In another variation, m is 2 to 7.

[0043] In another aspect, the naphthol derivative has the formula: Ia.1 , Ib.1 , Ic.1 , Ia.2 , Ib.2 , Ic.2 , IIa , IIb , IIc , IIa.1 , IIb.1 , IIc.1 , IIa.1.1 , IIb.1.1 , IIc.1.1 , IIa.1.2 , IIb.1.2 , IIc.1.2 , IIa.2 , IIb.2 , IIc.2 , IIa.3 , IIb.3 , IIc.3 , IIa.4 , IIb.4 , IIc.4 , IIa.5 , IIb.5 and IIc.5 In one variation, R 2 is H or CH3. In another variation, R 2 is CH3. In another aspect of the above naphthol derivative, n is 7. In another aspect, a pharmaceutical composition is provided, comprising a therapeutically effective amount of the above disclosed naphthol derivative or a mixture thereof; and a pharmaceutically acceptable excipient, wherein the composition is effective for treating a condition associated with vitamin K, selected from osteoporosis, atherosclerosis, calciphylaxis or tissue calcification, including soft tissue calcification.

[0044] In another variation of each of the above embodiments and aspects, n is 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14.

[0045] Treatment of COPD:

[0046] Chronic obstructive pulmonary disease (COPD) is a term used to describe a progressive lung disease that makes breathing difficult. The two main forms of COPD are emphysema and chronic bronchitis. In addition, elastin dissolution (proteolysis of elastin) is a key feature of COPD. Elastin dissolution can lead to loss of arterial flexibility and promote calcification of the intima-media membrane of blood vessels. Elastin dissolution has also been shown to be a strong predictor of mortality in COPD patients (Rabinovich et al., (2016) Circulating desmosine levels do not predict emphysema progression but are associated with cardiovascular risk and mortality in COPD, ERJ Express doi: 10.1183 / 13993003.01824-2015). MGP has been shown to inhibit the production of matrix metalloproteinases that promote elastin dissolution. Vitamin D may be a key determinant of the rate of elastin degradation, and low vitamin D levels may result in low MGP activity, insufficient to prevent elastin dissolution (Piscaer et al., (2017) Vitamin D deficiency: the linking pin between COPD and cardiovascular diseases? Respiratory Research (RESP.RES.) 18: 189). Without wishing to be bound by theory, as disclosed herein, enhancing the production of activated (carboxylated) MGP by administering vitamin K2 may be used to inhibit the deleterious effects of elastin dissolution on subjects with COPD, thereby preventing one or more symptoms of COPD or delaying the progression of one or more symptoms or reversing one or more symptoms. In addition, treating the degradation of elastin may be effective for treating Covid, such as Covid-19 and its variants. Therefore, the nanoparticle formulations disclosed herein may be administered for treating or preventing the degradation of elastin and diseases associated with the degradation of elastin.

[0047] In one embodiment, a method for treating, preventing, delaying the progression of, preventing and / or reversing tissue calcification or calciprotection in a mammal (or subject) is provided, the method comprising administering at least 0.1 mg of the above composition to the mammal daily to prevent tissue calcification, delay the progression of, and / or prevent tissue calcification, wherein the above composition is administered in the form of a pharmaceutical composition. In another embodiment, the present application discloses a method for increasing the tissue concentration of naphthol, which is a cofactor of gamma-glutamate carboxylase (GGCX), which is used to catalyze the carboxylation of vitamin K-dependent proteins, and is related to the treatment or prevention of osteoporosis, arteriosclerosis, calciprotection or tissue calcification in patients in need thereof, the method comprising administering a therapeutically effective amount of a naphthol derivative, or a pharmaceutical composition comprising an effective amount of the above naphthol derivative or a mixture thereof.

[0048] On the other hand, a method for treating a disease in a mammal is provided, wherein the disease is selected from the group consisting of: neurodegenerative diseases, retinopathy, rheumatoid polyarthritis, atherosclerosis, amyotrophic lateral sclerosis, cerebral ischemia, cataracts, systemic infection, pathologies associated with skin aging and tissue aging, pathologies associated with mitochondrial dysfunction, cachexia associated with malnutrition, wherein the treatment is associated with an extension of the life span of the mammal, the method comprising administering a therapeutically effective amount of a compound or composition comprising the above-mentioned naphthol derivative or a mixture thereof.

[0049] In another aspect, a method for treating a mammal suffering from a disease is provided, wherein the disease is selected from the group consisting of vitamin K deficiency, osteoporosis, proliferative diseases and cardiovascular diseases, the method comprising administering to the mammal a therapeutically effective amount of the above naphthol derivative or a mixture thereof. In another aspect of the method, the proliferative disease is selected from the group consisting of cancer, leukemia and inflammatory diseases.

[0050] On the other hand, a method for treating or preventing osteoporosis and / or osteopenia is provided, the method comprising administering a therapeutically effective amount of a composition comprising the above-mentioned naphthol derivative or a mixture thereof to a patient in need of treatment. On the other hand, a method for treating, preventing, delaying the progression of, preventing and / or reversing calciphylaxis in a mammal in need is provided, the method comprising administering a therapeutically effective amount of a composition comprising a substantially pure naphthol derivative or a mixture thereof as disclosed above and a pharmaceutically acceptable excipient to the mammal to prevent, delay the progression of, prevent or reverse calciphylaxis. In another aspect of the method, the mammal suffers from distal calciphylaxis and / or central calciphylaxis. In another aspect of the method, the mammal suffers from diabetes, chronic kidney disease or end-stage renal disease. In another aspect of the method, the mammal suffers from stage 3, stage 4 or stage 5 chronic kidney disease. In another aspect of the above, the mammal is undergoing hemodialysis. In another aspect, the mammal is receiving non-warfarin-based anticoagulant therapy. On the other hand, the anticoagulant therapy is an oral anticoagulant therapy. On the other hand, the anticoagulant therapy includes a factor Xa activity inhibitor selected from apixaban, rivaroxaban, betrixaban, edoxaban, otamixaban, letaxaban, eribaxaban or fondaparinux; or a factor IIa activity inhibitor selected from dabigatran or argatroban.

[0051] In another aspect of the above method, the mammal suffers from chronic obstructive pulmonary disease (COPD). On the other hand, the mammal suffers from skin lesions associated with calciphylaxis. In another aspect of the method, the total surface area of ​​the skin lesions is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% by applying the composition. In another aspect of the method, relative to the serum T50 value of the mammal before applying the compound disclosed above or its mixture, the mammal is administered with a compound as disclosed above or its mixture so that the serum T50 value of the mammal is increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. On the other hand, the administration of the above compound or its mixture increases the ratio of carboxylated vitamin K-dependent proteins to non-carboxylated vitamin K-dependent proteins in the plasma of the mammal, and the ratio after the administration of the composition is greater than the ratio before the administration of the composition.

[0052] On the other hand, a method for treating, preventing, delaying the progression of, preventing and / or reversing tissue calcification in a prediabetic mammal (or subject) suffering from diabetes, chronic kidney disease or a combination thereof and in need thereof is provided, the method comprising administering to the mammal at least 50 mg of a compound as disclosed above or a mixture thereof per day to prevent tissue calcification, delay the progression of tissue calcification and / or prevent tissue calcification, wherein the compound as disclosed above or a mixture thereof is administered in the form of a pharmaceutical composition. In one aspect of the method, the mammal suffers from diabetes. In another aspect of the method, the mammal suffers from type II diabetes; or the mammal has been diagnosed with prediabetes. In another aspect, the mammal suffers from chronic kidney disease; or the mammal suffers from stage 4 or stage 5 chronic kidney disease / end-stage renal disease. In another aspect of the method, the mammal is undergoing hemodialysis; or the mammal is receiving a non-warfarin-based anticoagulant therapy. In one variation, the anticoagulant therapy is oral anticoagulant therapy. In another aspect of the method, the anticoagulant therapy comprises an inhibitor of factor Xa activity selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban, or fondaparinux; or an inhibitor of factor IIa activity selected from dabigatran or argatroban.

[0053] In another aspect, a method for treating, preventing, slowing the progression of, preventing and / or reversing tissue calcification in a mammal undergoing hemodialysis and in need thereof is provided, the method comprising administering at least 5 mg of any one of the above compounds or a mixture thereof to the mammal daily, thereby preventing, slowing the progression of, preventing and / or reversing tissue calcification, wherein the above compounds or mixtures thereof are administered in the form of a pharmaceutical composition. In another aspect of the method, the mammal suffers from diabetes.

[0054] In another aspect, a method for increasing the bioavailability and serum half-life of at least one of menaquinone-7 / menaquinol-7, menaquinone-8 / menaquinol-8, menaquinone-9 / menaquinol-9, or menaquinone-10 / menaquinol-10 is provided. The method comprises administering or supplementing the above-mentioned compound or a composition thereof to a mammal in need thereof, wherein the method increases at least one of the following bioavailability and serum half-life by 5% when compared to administration or supplementation using the corresponding menaquinone-7, menaquinone-8, menaquinone-9 or menaquinone-10: menaquinone-7 / menaquinol-7, menaquinone-8 / menaquinol-8, menaquinone-9 / menaquinol-9 or menaquinone-10 / menaquinol-10.

[0055] On the other hand, a method for treating at least one of a wound or pain and inflammation area caused by calciphylaxis or tissue calcification in a subject in need is provided, the method comprising applying a dermal structure or a transdermal patch locally to the wound or pain and inflammation area of ​​the subject, wherein the dermal structure or the transdermal patch comprises any one of the above-mentioned compounds to treat the wound, pain and inflammation area. In a variation of the method, the local application of the dermal structure or the transdermal patch is performed before, in conjunction with (or simultaneously with) or after the above-mentioned method.

[0056] In another embodiment, a method for treating cancer is provided, wherein the cancer is selected from the group consisting of melanoma, lung cancer, breast cancer, leukemia, neuroblastoma, glioblastoma, cervical cancer, liver cancer (hepatocellular carcinoma, HCC), colorectal cancer, pancreatic cancer, bladder cancer, kidney cancer, prostate cancer, ovarian cancer and head and neck cancer.

[0057] In another embodiment, a pharmaceutical composition is provided, comprising a therapeutically effective amount of a formula according to any one of the above embodiments, aspects and variations. I , II and III A naphthol derivative or a mixture of naphthol derivatives thereof; and a pharmaceutically acceptable excipient, wherein the composition is effective in treating a condition associated with vitamin K selected from osteoporosis, atherosclerosis, calciphylaxis or tissue calcification. As used herein, unless otherwise specifically stated, the formula I , II and IIIThe naphthol compounds or derivatives include naphthol derivatives (or compounds) as disclosed herein, including the formula Ia.1 , Ib.1 , Ic.1 , Ia.2 , Ib.2 , Ic.2 , IIa , IIb , IIc , IIa.1 , IIb.1 , IIc.1 , IIa.1.1 , IIb.1.1 , IIc.1.1 , IIa.1.2 , IIb.1.2 , IIc.1.2 , IIa.2 , IIb.2 , IIc.2 , IIa.3 , IIb.3 , IIc.3 , IIa.4 , IIb.4 , IIc.4 , IIa.5 , IIb.5 and IIc.5 ;as well as III . In one variation of the pharmaceutical composition, the composition comprises sunflower oil. In another variation of the pharmaceutical composition, the composition comprises an oil selected from the group consisting of sunflower oil, corn oil, vegetable oil (such as jojoba oil) or a combination thereof. In another variation, the composition comprises an oil selected from the group consisting of canola oil, coconut oil, cottonseed oil, olive oil, palm oil, peanut oil, rapeseed oil, safflower oil, sesame oil, soybean oil, heliotrope oil, almond oil, cashew oil, hazelnut oil, walnut oil, pecan oil, pine nut oil, pistachio oil, castor oil and a combination thereof.

[0058] In another embodiment, a method is provided for increasing the tissue concentration of naphthol as a cofactor for gamma-glutamate carboxylase (GGCX), wherein GGCX is used to catalyze the carboxylation of vitamin K-dependent proteins associated with the treatment or prevention of osteoporosis, atherosclerosis, calciphylaxis, or tissue calcification in a patient in need thereof, the method comprising administering a therapeutically effective amount of a naphthol derivative, or a pharmaceutical composition comprising an effective amount of a naphthol derivative of any one of the above-mentioned naphthol derivatives, or a mixture thereof.

[0059] In one variation, the method increases tissue concentrations of naphthol by at least 20%, 30%, 40%, 50%, 60%, 70% or more when compared to administration of the corresponding menadione. In another variation, the method increases tissue concentrations of naphthol by at least 40% or 50% when compared to administration of the corresponding menadione. In another variation, the method increases tissue concentrations of naphthol by at least 20%, 30%, 40%, 50%, 60%, 70% or more when compared to administration of the corresponding menadione, and provides optimal benefit at the tissue level, such as at least one of skin or dermal tissue, mitral valve, aortic valve, and blood vessels.

[0060] In another embodiment, a method for treating a disease in a mammal is provided, wherein the disease is selected from the group consisting of: neurodegenerative diseases, retinopathy, rheumatoid polyarthritis, atherosclerosis, amyotrophic lateral sclerosis, cerebral ischemia, cataracts, systemic infection, pathologies associated with skin aging and tissue aging, pathologies associated with mitochondrial dysfunction, cachexia associated with malnutrition, wherein the treatment is associated with an extension of the lifespan of the mammal, the method comprising administering a therapeutically effective amount of a compound or composition comprising a naphthol derivative according to any one of the above embodiments, aspects and variations or a mixture of the above naphthol derivatives.

[0061] In another embodiment, a method for treating a mammal suffering from a disease is provided, wherein the disease is selected from the group consisting of vitamin K deficiency, osteoporosis, proliferative diseases and cardiovascular diseases, the method comprising administering to the mammal a therapeutically effective amount of any one of the above naphthol derivatives or a mixture thereof. In one aspect of the method, the proliferative disease is selected from the group consisting of cancer, leukemia and inflammatory diseases.

[0062] In another embodiment, a method for treating or preventing osteoporosis and / or bone mass loss is provided, the method comprising administering to a patient in need of treatment a therapeutically effective amount of a composition comprising a naphthol derivative according to any one of the above embodiments, aspects and variations or a mixture of the above naphthol derivatives.

[0063] In another embodiment, a method for treating, preventing, delaying the progression of, preventing and / or reversing calciphylaxis in a mammal in need thereof is provided, the method comprising administering to the mammal a therapeutically effective amount of a composition comprising a substantially pure calciphylaxis derivative of any one of the above-mentioned calciphylaxis derivatives or a mixture thereof and a pharmaceutically acceptable excipient to prevent, delay the progression of, prevent or reverse calciphylaxis. In one aspect of the above method, the mammal suffers from distal calciphylaxis and / or central calciphylaxis. In another aspect of the method, the mammal suffers from diabetes, chronic kidney disease or end-stage renal disease. In another aspect of the method, the mammal suffers from stage 3, stage 4 or stage 5 chronic kidney disease. In another aspect of the method, the mammal is undergoing hemodialysis. In another aspect of the method, the mammal is receiving non-warfarin-based anticoagulant therapy. In another aspect of the method, the anticoagulant therapy is oral anticoagulant therapy. In another aspect of the method, the anticoagulant therapy comprises an inhibitor of factor Xa activity selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban, or fondaparinux; or an inhibitor of factor IIa activity selected from dabigatran or argatroban.

[0064] In another aspect of the above method, the mammal suffers from chronic obstructive pulmonary disease (COPD). In another aspect of the method, the mammal suffers from calciphylaxis-related skin lesions. In another aspect of the method, the application of the composition reduces the total surface area of ​​the skin lesions by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. As used herein, the method reduces the surface area of ​​skin lesions, which helps the healing of the skin or the healing of wounds; or the method thus enhances or accelerates wound healing. Therefore, this method is generally applicable to wound healing, such as skin burns (or burns), ulcers, and wounds generally caused by trauma, skin cancer, infection and cosmetic surgery. In addition, as disclosed below, the above method can be used in combination with or simultaneously with the topical application of the composition to further promote wound healing.

[0065] In another aspect of the above method, administration of any one of the naphthol derivatives (or compounds) or a mixture thereof according to the above embodiments, aspects and variations to the mammal increases the serum T50 value of the mammal by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the serum T50 value of the mammal before administration of the compounds or mixtures thereof disclosed above.

[0066] In another aspect of the method, administration of a naphthol compound according to any of the above embodiments, aspects and variations increases the ratio of carboxylated vitamin K-dependent proteins to non-carboxylated vitamin K-dependent proteins in the plasma of the mammal, with the ratio after administration of the composition being greater than the ratio before administration of the composition.

[0067] In another embodiment, a method for treating, preventing, delaying the progression of, preventing and / or reversing tissue calcification in a prediabetic mammal (or subject) with diabetes, chronic kidney disease or a combination thereof in need is provided, the method comprising administering at least 50 mg of a compound according to any one of the above embodiments, aspects and variations to the mammal daily to prevent tissue calcification, delay the progression of, and / or prevent tissue calcification, wherein the above compound or a mixture thereof is administered in the form of a pharmaceutical composition. In another aspect of the method, the mammal suffers from diabetes. In another aspect of the method, the mammal suffers from type II diabetes.

[0068] In another aspect of the above method, the mammal has been diagnosed with prediabetes. In another aspect of the method, the mammal suffers from chronic kidney disease. In another aspect of the method, the mammal suffers from stage 4 or stage 5 chronic kidney disease / end-stage renal disease. In another aspect of the method, the mammal is undergoing hemodialysis. In another aspect of the method, the mammal is receiving non-warfarin-based anticoagulant therapy. In another aspect of the method, the anticoagulant therapy is oral anticoagulant therapy. On the other hand, the anticoagulant therapy includes a factor Xa activity inhibitor selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irishaban or fondaparinux sodium; or a factor IIa activity inhibitor selected from dabigatran or argatroban.

[0069] In another embodiment, a method for treating, preventing, slowing the progression of, preventing and / or reversing tissue calcification in a mammal undergoing hemodialysis and in need thereof is provided, the method comprising administering to the mammal at least 5 mg of a compound according to any one of the above embodiments, aspects and variations per day, thereby preventing, slowing the progression of, preventing and / or reversing tissue calcification, wherein the compound according to any one of the above embodiments, aspects and variations or a mixture thereof is administered in the form of a pharmaceutical composition. In another aspect, the mammal suffers from diabetes.

[0070] In another embodiment, a method is provided for increasing at least one of the bioavailability and serum half-life of: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9 or menaquinone-10 / naphthol-10, the method comprising administering or supplementing a compound or composition according to any one of the above embodiments, aspects and variations to a mammal in need thereof, wherein the method increases at least one of the bioavailability and the serum half-life of: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9 or menaquinone-10 / naphthol-10 by 5% when compared to administration or supplementation using menaquinone-7, menaquinone-8, menaquinone-9 or menaquinone-10.

[0071] Treatment of Alzheimer's Disease, Apoptosis and Cancer:

[0072] Alzheimer's disease (AD) is a devastating neurodegenerative disorder. Its sporadic form affects the elderly population (with a sharp increase in incidence in those aged >75 years), and in addition, there are various familial forms that develop in the fourth or fifth decade of life. AD is characterized by the presence of extracellular senile plaques and intracellular neurofibrillary tangles in the brain of patients. The core component of senile plaques is a small, 4kDa amyloid peptide, which is produced by proteolytic processing of the large transmembrane protein amyloid precursor protein (APP). Cleavage of APP by β-secretase (BACE-1) releases a soluble APP-β fragment, while the C-terminus of 99 amino acids in length remains tethered to the membrane. This C-terminal fragment is then proteolytically processed by γ-secretase (a membrane multienzyme complex) to produce amyloid peptides of varying lengths, primarily 40 and 42 amino acids in length (Hardy J. et al. (2002) Science; 297(5580):353-356). In one embodiment, the disease or condition treated using the compositions as disclosed herein is mediated by BACE-1, BACE-2 or cathepsin D activity, such as Alzheimer's disease, mild cognitive impairment, impaired glucose tolerance or type 2 diabetes. In one variation, the disclosed methods can be used to treat brain calcification.

[0073] It has been shown that beta-amyloid protein (Aβ) causes neuronal death by promoting apoptosis and also by direct toxicity. Hadipour, E et al., Vitamin K2 protects PC12 cells from Aβ (1-42) and H2O2-induced apoptosis via P38 MAP Kinase Pathway. "Nutr. Neurosci. " 2020, 23, 343-352. Neurotoxicity occurs through a variety of mechanisms, including the destruction of calcium homeostasis, oxidative stress and mitochondrial dysfunction. In PC12 cells derived from rat pheochromocytoma, vitamin K2 can prevent neuronal death caused by Aβ (1-42) (the most neurotoxic form of Aβ). The experiments also showed that cells pretreated with vitamin K2 exhibited significantly reduced apoptosis when the cells were exposed to hydrogen peroxide (H2O2) or Aβ(1-42). Pretreatment with vitamin K2 also reduced the amount of apoptosis signaling proteins, including a lower Bax / Bcl-2 ratio, reduced the presence of reactive oxygen species (ROS), and increased the amount of glutathione, which acts as an antioxidant. Yagami, T. Gas6 Rescues Cortical Neurons from AmyloidβProtein-Induced Apoptosis Neuropharmacology 2002, 43, 1289-1296. The researchers determined that inactivation of the p38 MAP kinase pathway is a mechanism for the potential protective effect of VK2 in Alzheimer's disease (AD).

[0074] The researchers observed that increased concentrations of vitamin K2 (VK2) prolonged cell survival, likely due to its protective effect against Aβ-induced neuronal death. It was also noted that this effect was reversible after the addition of warfarin, which prevented vitamin K-dependent carboxylation. VK2 was observed to reduce the number of ROS in a dose-dependent manner, and at a concentration of 10 mcmol / L, the activity of caspase-3, an enzyme that mediates Aβ-induced apoptosis, was reduced by 2.5-fold. The authors also found that Gas6 plays a role in VK2's protection against Aβ cytotoxicity; this was confirmed based on measured Ca(2+) influx, chromatin condensation, and DNA fragmentation as markers of Aβ neurotoxicity and apoptosis in rat embryonic neuronal cell cultures. Gas6 inhibited Ca(2+) influx in a dose-dependent manner and significantly reduced the amount of chromatin condensation and DNA fragmentation caused by Aβ. Therefore, there is a clear correlation between the antioxidant properties of vitamin K2 and its antiapoptotic properties, which are related to the process of programmed cell death as a potent mechanism relevant to cancer therapy.

[0075] Targeting apoptosis is also effective against all types of cancer, as apoptosis evasion is a hallmark of cancer. Apoptosis is also not specific to the cause or type of cancer. See Villa-Pulgarín JA et al., Mitochondrial and lipid raft localization of F o Mitochondria and lipid raft-located F1-ATP synthase as the main therapeutic target for the antileishmanial and anticancer activities of ether lipid edilfosine o F1-ATP synthase as major therapeutic targets in the antileishmanial and anticancer activities of ether lipid) "PLoS Negl. Trop. Dis." 2017;11:e0005805.doi:10.1371 / journal.pntd.0005805; Elmore S. Apoptosis: A review of programmed cell death "Toxicol. Pathol. 2007;35:495-516.doi:10.1080 / 01926230701320337.

[0076] Vitamin K2 is also used in clinical applications to supplement cancer treatment. XV, F. et al., Research Progress on the anticancer effects of vitamin K2 (Review). Oncol. Lett. 2018, 15, 8926-8934. In addition, vitamin K2 supplementation has been found to prevent the growth and metastasis of multiple cancer lines. See Xia, J. et al., The role of PKC isoforms in the inhibition of NF-κB activation by vitamin K2 in human hepatocellular carcinoma cells. J. Nutr. Biochem. 2012m 23, 1668-1675; Showalter, SL et al., Naturally occurring K vitamins inhibit pancreatic cancer cell survival through a caspase-dependent pathway. J. Gastroenterol. Hepatol. 2010, 25, 738-744. Enomoto, M. et al., Vitamin K2-induced cell growth inhibition via autophagy formation in cholangiocellular carcinoma cell lines. International Journal of Molecular Medicine 2007, 20, 801-808; Jinghe, X., Vitamin K and hepatocellular carcinoma: The basic and clinic. World J. Clin. Cases 2015, 3, 757-764.Sada, E. et al., Vitamin K2 modulates differentiation and apoptosis of both myeloid and erythroid lineages. Eur. J. Haematol. 2010, 85, 538-548; Yaguchi, M. et al., Vitamin K2 and its derivatives induce apoptosis in leukemia cells and enhance the effect of all-trans retinoic acid. Leukemia 1997, 11, 779-787.

[0077] In fact, it has been determined that the intake of MK-4 and MK5 to MK-9 is negatively correlated with cancer mortality. Specifically, in a male cohort study, the incidence of advanced prostate cancer was negatively correlated with the intake of MK (especially MK-5 to MK-9). Nimptsch, K. et al. (2008) Dietary intake of vitamin K and risk of prostate cancer in the Heidelberg cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC-Heidelberg) "American Journal of Clinical Nutrition (Am. J. Clin. Nutr.)" 87, 985-992. Nimptsch, K. et al. (2010) Dietary vitamin K intake in relation to cancer incidence and mortality: results from the Heidelberg cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC-Heidelberg). Am J Clin Nutr 91, 1348-1358. In a nested case-control follow-up study, incomplete carboxylation of osteocalcin, a marker of inadequate vitamin K status, was significantly higher in cases with advanced or high-grade prostate cancer compared with controls.Nimptsch, K. et al. (2009) Serum undercarboxylated osteocalcin as biomarker of vitamin K intake and risk of prostate cancer: a nested case-control study in the Heidelberg cohort of the European Prospective Investigation into Cancer and Nutrition. Cancer Epidemiol. Biomark. Prev. 18, 49-56. Wang et al. also reported a positive association between MK intake and breast cancer incidence and mortality risk, especially for luminal, triple-negative, and early-stage disease. Wang, K. et al. (2021) Vitamin K intake and breast cancer incidence and mortality: results from a prospective cohort study. Clin. Nutr. 40, 3370-3378. Therefore, in one variation, a method for treating cancer (such as prostate cancer) by administering a therapeutically effective amount of the disclosed composition is provided to control or reduce high levels of incomplete carboxylation of osteocalcin.

[0078] Stability of compounds in SGF, FaSSIF and FeSSIF:

[0079] The stability of the disclosed compounds (such as the disclosed compounds tested) can be determined in simulated gastric fluid (SGF, pH-1.2), fasted simulated intestinal fluid (FaSSIF, pH-6.5) and fed simulated intestinal fluid (FeSSIF, pH-5) at 0 minutes, 30 minutes, 60 minutes, 120 minutes and 240 minutes. The % disappearance of the compound over time can be calculated by HPLC / LCMS analysis, comparing the peak area of ​​the analyte relative to the '0' minute. As disclosed herein, the formation of menaquinones, such as menaquinone-7 (i.e., MK-7), was observed by the tested compounds.

[0080] The disclosed formula tested I , II orIII The stability of the compounds was recorded for SGF (pH 1.2); FaSSIF (pH 6.5) and FeSSIF (pH 5.0) at time intervals of t = 0 min, 30 min, 60 min, 120 min and 240 min, and the % of compound remaining compared to 0 min was recorded.

[0081] The disclosed formula tested I , II or III The stability of the compounds was recorded for SGF (pH 1.2); FaSSIF (pH 6.5) and FeSSIF (pH 5.0) at time intervals of t = 0 min, 30 min, 60 min, 120 min and 240 min, and the formation of menaquinone-7 was recorded as fold change compared to 0 min. I , II or III The stability of the compounds can be determined in simulated gastric fluid (SGF, pH-1.20), fasted state simulated intestinal fluid (FaSSIF, pH-6.5) and fed state simulated intestinal fluid (FeSSIF, pH-5) at 0, 30, 60, 120 and 240 minutes. The % disappearance of the compound over time is calculated by comparing the peak area of ​​the analyte relative to that at '0' minute by HPLC / LCMS analysis. The formation of menaquinone-7 from the test material was observed. Stability in human and rat plasma: The disclosed compounds are stable over time.

[0082] The disclosed formula tested was determined in human and rat plasma (K2EDTA) at 0, 15, 30, 60 and 120 minutes. I , II or III Stability of Compounds. The % disappearance of the compound over time compared to 0 min was calculated by HPLC / LC-MS-MS analysis. The formation of menaquinone-7 by the tested compounds was recorded at time intervals of t = 0 min, 30 min, 60 min and 120 min.

[0083] The disclosed formula tested was determined in human and rat plasma (K2EDTA) at 0, 15, 30, 60 and 120 minutes. I , II or III Stability of compounds. The formation of menaquinone-7 based on fold change compared to 0 min was calculated by HPLC / LC-MS-MS analysis. The % remaining of the tested compound was observed at time intervals of t=0 min, 30 min, 60 min and 120 min.

[0084] The disclosed formula tested was determined in human and rat plasma (K2EDTA) at 0, 15, 30, 60 and 120 minutes. I , II or III The stability of the tested compounds was analyzed by HPLC / LC-MS-MS and the formation of menaquinone-7 was calculated based on the fold change compared to 0 min and observed at time intervals of t=0 min, 30 min, 60 min and 120 min.

[0085] The disclosed formula tested was determined in human and rat plasma (K2EDTA) at 0, 15, 30, 60 and 120 minutes. I , II or III Stability of Compounds. The % disappearance of the compound over time compared to 0 min was calculated by HPLC / LC-MS-MS analysis. The formation of menaquinone-7 by the tested compounds was recorded.

[0086] In vivo testing:

[0087] Male Sprague Dawley rats of 8-10 weeks of age and weighing approximately 202-223 grams were used for in vivo testing. The animals were fasted overnight and had free access to water. The animals were divided into 4 groups G1 to G4, each containing 3 rats. To determine the bioavailability, the test substances were dissolved in corn oil to obtain a uniform formulation. The animals were administered the test substances by gavage, respectively, G1 animals received a dose of 1088 μg / kg body weight of the compound Ia.1 , m=9, n=7 (G1), G2 received 1073 μg / kg body weight of the compound Ia.1 , m = 9, n = 9 (G2), G3 and G4 received 1000 μg / kg body weight of menaquinone-7 (G3) and menaquinone-9 (G4). Ia.1 (n=7)(G1) and Ia.1 Blood samples were collected at different time points during the 48 hours after dosing.

[0088] Blood samples were collected and transferred to tubes containing K2EDTA and immediately placed on ice for plasma preparation. Plasma was prepared by centrifugation at 3500 g for 10 minutes and aliquots were frozen at -80 °C until analysis. Quantification of analytes in plasma was determined by LC-MS-MS analysis. Plasma PK parameters were calculated using WinNonlin software.

[0089] In another embodiment, the present application discloses a pharmaceutical composition comprising a therapeutically effective amount of a naphthol derivative or a mixture thereof according to any one of the above embodiments and aspects; and a pharmaceutically acceptable excipient, wherein the composition can effectively treat a condition associated with vitamin K selected from osteoporosis, arteriosclerosis, calciphylaxis or tissue calcification.

[0090] In another embodiment, the present application discloses a method for increasing the tissue concentration of naphthol as a cofactor of gamma-glutamate carboxylase (GGCX), wherein GGCX is used to catalyze the carboxylation of vitamin K-dependent proteins associated with the treatment or prevention of osteoporosis, atherosclerosis, calciphylaxis or tissue calcification in patients in need thereof, the method comprising administering a therapeutically effective amount of a naphthol derivative, or a pharmaceutical composition comprising an effective amount of a naphthol derivative or a mixture thereof as disclosed above.

[0091] In one variation of the method, administration of the naphthol derivative overcomes oxidative block in patients with CKD and patients undergoing hemodialysis and provides maximum levels of naphthol and maximum benefit at the tissue level. In one variation, the tissue is skin or dermal tissue. In another variation, the tissue is at least the patient's mitral valve, the patient's aortic valve, and a blood vessel. In another variation of the method, the naphthol derivative is of the formula I , II or III In another variation, the method increases the tissue concentration of the corresponding menaquinone by at least 20%, 30%, 40%, 50%, 100%, 150%, 200%, 250%, 300%, 400%, or 500% or more when compared to administration of the corresponding menaquinone. In another variation, the method increases the tissue concentration of the corresponding menaquinone by at least 40% or 50% when compared to administration of the corresponding menaquinone. "Corresponding menaquinone" means, for example, that when the naphthol derivative is a MK-7 derivative, then the "corresponding menaquinone" will be MK-7; and when the naphthol derivative is a MK-10 derivative, then the corresponding menaquinone will be MK-10.

[0092] In another embodiment, the present application discloses a method for treating a disease in a mammal, the disease being selected from the group consisting of: neurodegenerative disease, retinopathy, rheumatoid polyarthritis, atherosclerosis, amyotrophic lateral sclerosis, cerebral ischemia, cataracts, systemic infection, pathology associated with skin aging and tissue aging, pathology associated with mitochondrial dysfunction, cachexia associated with malnutrition, wherein the treatment is associated with an extension of the life span of the mammal, the method comprising administering a therapeutically effective amount of a compound or composition comprising the formula I ,II or III (including all disclosed compounds) naphthol derivatives or mixtures thereof. As described above, the formula I , II or III The naphthol compounds or derivatives include those disclosed herein and those having the formula Ia , Ib , Ic , Ia.1 , Ib.1 , Ic.1 , IIA , IIb and IIc naphthol derivatives (or compounds); and those compounds as disclosed herein.

[0093] On the other hand, the present application discloses a method for treating a mammal with a disease, the disease being selected from the group consisting of: vitamin K deficiency, osteoporosis, proliferative diseases and cardiovascular disease, the method comprising administering to the mammal a therapeutically effective amount of a naphthol derivative as disclosed above or a mixture thereof. In another aspect of the method, the proliferative disease is selected from the group consisting of cancer, leukemia and inflammatory diseases. In one variation of the method, cancer is melanoma and lung cancer. In another variation, cancer is selected from the group consisting of: leukemia, neuroblastoma, glioblastoma, cervical cancer, liver cancer (hepatocellular carcinoma, HCC), colorectal cancer, pancreatic cancer, bladder cancer, kidney cancer and melanoma. In another aspect of the method, cancer is selected from the group consisting of: lung cancer, breast cancer, prostate cancer, ovarian cancer and head and neck cancer.

[0094] In another aspect, the present application discloses a method for treating or preventing osteoporosis and / or osteopenia, the method comprising administering to a patient in need of treatment a therapeutically effective amount of a composition comprising the formula I , II or III A naphthol derivative or a mixture thereof.

[0095] In another aspect, the present application discloses a method of treating, preventing, slowing the progression of, arresting and / or reversing calciphylaxis in a mammal in need thereof, the method comprising administering to the mammal a therapeutically effective amount of a composition comprising a substantially pure compound of formula I , II or IIIa naphthol derivative or a mixture thereof and a pharmaceutically acceptable excipient to prevent the calciphylaxis, delay the progression of the calciphylaxis, prevent or reverse the calciphylaxis. In another aspect of the method, the mammal suffers from distal calciphylaxis and / or central calciphylaxis. In another aspect of the method, the mammal suffers from diabetes, chronic kidney disease or end-stage renal disease. In another aspect of the method, the mammal suffers from stage 3, stage 4 or stage 5 chronic kidney disease. In another aspect of the method, the mammal is undergoing hemodialysis. In another aspect of the method, the mammal is receiving non-warfarin-based anticoagulant therapy. In another aspect of the method, the anticoagulant therapy is oral anticoagulant therapy. In another aspect of the method, the anticoagulant therapy includes a factor Xa activity inhibitor selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irixaban or fondaparinux sodium; or a factor IIa activity inhibitor selected from dabigatran or argatroban.

[0096] In another aspect of the above method, the mammal has chronic obstructive pulmonary disease (COPD). In another aspect of the method, the mammal has calciphylaxis-related skin lesions. In another aspect, administration of the composition reduces the total surface area of ​​the skin lesions by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In another aspect of the method, administration of the composition to the mammal reduces the total surface area of ​​the skin lesions by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, relative to the serum T50 value of the mammal before administration of the above compound or mixture thereof. I , II or III The compound or mixture thereof increases the serum T50 value of the mammal by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. In another aspect of the method, administering the formula I , II or III The compound or mixture thereof increases the ratio of carboxylated vitamin K-dependent protein to non-carboxylated vitamin K-dependent protein in the plasma of the mammal, wherein the ratio after administration of the composition is greater than the ratio before administration of the composition.

[0097] In another embodiment, a method for treating, preventing, slowing the progression of, preventing and / or reversing tissue calcification in a prediabetic mammal (or subject) having diabetes, chronic kidney disease or a combination thereof in need thereof is provided, the method comprising administering to the mammal at least 50 mg of the formula I , II or IIIA compound or a mixture thereof to prevent tissue calcification, delay the progression of tissue calcification and / or stop the tissue calcification, wherein the above-mentioned compound or the mixture thereof is administered in the form of a pharmaceutical composition.

[0098] In each of the above methods, the specification also discloses the use of the compound or its composition for treating the medical condition or disease; and the specification discloses the use of the compound in the preparation of a formulation or medicament for treating the disclosed medical condition or disease.

[0099] In one variation, the naphthol derivative is administered at a dose of at least 1 mg, 3 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, or 50 mg. In another variation, the naphthol derivative is administered at a dose of at least 100 mg, 150 mg, 200 mg, 25 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg. In another variation, the naphthol derivative is administered at a dose of at least 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1,000 mg or more.

[0100] In one aspect of the above method, the mammal suffers from diabetes. In another aspect, the mammal suffers from type II diabetes. In another aspect, the mammal has been diagnosed with prediabetes. In another aspect of the above method, the mammal suffers from chronic kidney disease. In another aspect of the method, the mammal suffers from stage 4 or stage 5 chronic kidney disease / end-stage renal disease. In another aspect of the method, the mammal is undergoing hemodialysis. In another aspect of the method, the mammal is receiving non-warfarin-based anticoagulant therapy. In another aspect of the method, the anticoagulant therapy is oral anticoagulant therapy. In another aspect of the method, the anticoagulant therapy includes a factor Xa activity inhibitor selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irishaban or fondaparinux sodium; or a factor IIa activity inhibitor selected from dabigatran or argatroban.

[0101] In another embodiment, the present application discloses a method for treating, preventing, slowing the progression of, arresting and / or reversing tissue calcification in a mammal undergoing hemodialysis and in need thereof, the method comprising administering to the mammal at least 5 mg of a formula I , II or III A compound or a mixture thereof, thereby preventing tissue calcification, delaying the progression of tissue calcification, arresting and / or reversing the tissue calcification, wherein the above-mentioned compound or mixture thereof is administered in the form of a pharmaceutical composition.

[0102] In one variation, the naphthol derivative is administered at a dosage of at least 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg or 50 mg. In another variation, the naphthol derivative is administered at a dosage of at least 100 mg, 150 mg, 200 mg, 25 mg, 300 mg, 350 mg, 400 mg, 450 mg or 500 mg. In another variation, the naphthol derivative is administered at a dosage of at least 500 mg, 600 mg, 700 mg, 800 mg, 900 mg or 1,000 mg or more. In another aspect of the above method, the mammal suffers from diabetes.

[0103] On the other hand, the present application discloses a pharmaceutical composition comprising a therapeutically effective amount of a naphthol derivative as disclosed above (or also referred to as a "compound" or a "disclosed compound") or a mixture thereof; and a pharmaceutically acceptable excipient, wherein the composition can effectively treat a condition associated with vitamin K selected from osteoporosis and arteriosclerosis.

[0104] On the other hand, the present application discloses a method for treating a disease in a mammal, wherein the disease is selected from the group consisting of: neurodegenerative diseases, retinopathy, rheumatoid polyarthritis, atherosclerosis, amyotrophic lateral sclerosis, cerebral ischemia, cataracts, systemic infection, pathologies associated with skin aging and tissue aging, pathologies associated with mitochondrial dysfunction, cachexia associated with malnutrition, wherein the treatment is associated with an extension of the lifespan of the mammal, the method comprising administering a therapeutically effective amount of a compound or composition comprising a naphthol compound or a mixture thereof as disclosed above.

[0105] In another embodiment, a method for treating a mammal suffering from a disease selected from the group consisting of vitamin K deficiency, osteoporosis, proliferative diseases and cardiovascular disease is provided, the method comprising administering to the mammal a therapeutically effective amount of a compound as disclosed herein or a mixture thereof. In another aspect of the method, the proliferative disease is selected from the group consisting of cancer, leukemia and inflammatory diseases.

[0106] In another embodiment, a method for treating or preventing osteoporosis and / or osteopenia is provided, the method comprising administering to a patient in need of treatment a therapeutically effective amount of a composition comprising a compound as disclosed above or a mixture thereof. The disclosed methods for administering compounds and compositions or combinations thereof can be used to treat or reduce vascular calcification, increase bone mineral density, and treat, reduce or prevent bone diseases, such as patients with CKD.

[0107] In another embodiment, a method for treating, preventing, slowing the progression of, preventing and / or reversing calciphylaxis in a mammal in need thereof is provided, the method comprising administering to the mammal a therapeutically effective amount of a composition comprising a substantially pure naphthol compound as disclosed herein and a pharmaceutically acceptable excipient to prevent, slow the progression of, prevent or reverse calciphylaxis. In one aspect of the method, the mammal suffers from distal calciphylaxis and / or central calciphylaxis. In another aspect, the mammal suffers from diabetes, chronic kidney disease or end-stage renal disease. In another aspect, the mammal suffers from stage 3, stage 4 or stage 5 chronic kidney disease. In another aspect of the method, the mammal is undergoing hemodialysis. In another aspect, the mammal is receiving non-warfarin-based anticoagulant therapy.

[0108] In another aspect of the above method, the anticoagulant therapy is oral anticoagulant therapy. On the other hand, the anticoagulant therapy includes a factor Xa activity inhibitor selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban or fondaparinux; or a factor IIa activity inhibitor selected from dabigatran or argatroban. On the other hand, the mammal suffers from chronic obstructive pulmonary disease (COPD). On the other hand, the mammal suffers from skin lesions associated with calciphylaxis. On the other hand of the method, the total surface area of ​​the skin lesions is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% by applying the composition. In another aspect of the method, administration of a substantially pure compound as disclosed herein to the mammal increases the serum T50 value of the mammal by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the serum T50 value of the mammal prior to administration of the disclosed compound. In another aspect, administration of the disclosed compound increases the ratio of carboxylated vitamin K-dependent protein to non-carboxylated vitamin K-dependent protein in the plasma of the mammal, the ratio after administration of the composition being greater than the ratio before administration of the composition. In one aspect of the method, the ratio of carboxylated vitamin K-dependent protein to non-carboxylated vitamin K-dependent protein in the plasma of the mammal increases by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% after administration of the composition relative to the ratio before administration.

[0109] In certain embodiments above, administration of the disclosed compounds reduces the amount of non-carboxylated vitamin K-dependent protein in the subject's plasma by, for example, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, relative to the amount prior to administration of the compound. In certain variations, the vitamin K-dependent protein is selected from matrix Gla protein (MGP), growth arrest-specific gene 6 (Gas-6) protein, PIVKA-II protein, osteocalcin, activated protein C, activated protein S, factor II, factor VII, factor IX, and factor X.

[0110] In certain variations of the above methods, administration of the compound increases the plasma levels of osteoprotegerin and fetuin A by, for example, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% relative to the plasma concentration of osteoprotegerin or fetuin A before administration of the compound. In other variations, administration of the compound reduces the plasma level of D-dimer or high-sensitivity C-reactive peptide (hs-CRP), e.g., by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, relative to the plasma concentration of D-dimer or high-sensitivity C-reactive peptide (hs-CRP) prior to administration of the compound.

[0111] In certain variations, the method may include administering to the subject about 10 mg to about 750 mg of the compound per day. In other variations, the method may include administering to the subject about 50 mg to about 750 mg of the compound per day. In other variations, the method may include administering to the subject about 20 mg to about 500 mg of the compound per day. In other variations, the method may include administering to the subject about 50 mg to about 500 mg of the compound per day. In certain variations, the method may include administering to the subject about 20 mg to about 250 mg of the compound per day. In other variations, the method may include administering to the subject about 5 mg to about 250 mg of the compound per day. In other variations, the method may include administering to the subject about 20 mg to about 100 mg of the compound per day. In other variations, the method may include administering to the subject about 50 mg to about 100 mg of the compound per day. In other variations, the methods may comprise administering to the subject from about 5 mg to about 75 mg of the compound per day, for example, administering to the subject 5, 10, 25, 50, 75, 100, 200, 300, 400, or 500 mg of the compound per day.

[0112] In certain variations, the compound is administered to the subject for at least 2 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 6 months, 1 year, or indefinitely as needed. If the subject is undergoing hemodialysis, the compound may be administered to the subject over a period of time that includes at least the duration of hemodialysis.

[0113] In another variation of the method for treating calciphylaxis, in addition to measuring the change / reduction in lesion size following administration of a disclosed compound before and after administration, the lesion of interest can be biopsied to determine tissue levels of PTH and evidence of changes in calcium and phosphate deposition in the cutaneous arterioles using von Kassa staining.

[0114] The presence of uremic oxidative blockade is determined by measuring: increased plasma lipid peroxidation, such as by detecting increased F2 isoprostanes, increased isolevuglandin-plasma protein adducts; increased protein and amino acid oxidation, such as by detecting oxidation of tyrosine residues, cysteine ​​or methionine residues, lysine oxidation and threonine oxidation in plasma proteins, thiol oxidation and carbonyl formation; reactive aldehyde formation, such as by detecting glyoxal, methylglyoxal, acrolein, glycolaldehyde and p-hydroxyphenylacetaldehyde; increased reactive carbonyl compounds, such as by measuring hydrazine formation following reaction with 2,4-dinitrophenylhydrazine; decreased plasma glutathione levels and glutathione peroxidase function; and an increased ratio of oxidized thiols to reduced thiols.

[0115] In another embodiment, a method of treating, preventing, slowing the progression of, preventing and / or reversing tissue calcification in a prediabetic mammal (or subject) having diabetes, chronic kidney disease or a combination thereof and in need thereof is provided, the method comprising administering to the mammal at least 0.2 mg or at least 2 mg of a substantially pure compound as disclosed herein per day to prevent, slow the progression of and / or prevent tissue calcification, wherein the compound is administered in the form of a pharmaceutical composition. In another aspect of the method, the mammal has diabetes. In another aspect, the mammal has type II diabetes; or the mammal has been diagnosed with prediabetes. In another aspect of the above method, the mammal has chronic kidney disease. In another aspect of the above method, the mammal has stage 4 or stage 5 chronic kidney disease / end-stage renal disease. In another aspect, the mammal is undergoing hemodialysis. In another aspect, the mammal is receiving a non-warfarin-based anticoagulant therapy. In another aspect, the anticoagulant therapy is oral anticoagulant therapy. In another aspect of the method, the anticoagulant therapy comprises an inhibitor of factor Xa activity selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban, or fondaparinux; or an inhibitor of factor IIa activity selected from dabigatran or argatroban.

[0116] In another embodiment, a method of treating, preventing, slowing the progression of, arresting and / or reversing tissue calcification in a mammal undergoing hemodialysis and in need thereof is provided, the method comprising administering to the mammal at least 0.2 mg or at least 2 mg per day of a substantially pure compound as disclosed herein, thereby preventing, slowing the progression of, arresting and / or reversing tissue calcification, wherein the disclosed compound is administered in the form of a pharmaceutical composition. In another aspect, the mammal suffers from diabetes.

[0117] Vitamin K metabolism: Development of vascular and soft tissue calcifications following failure of regeneration of the reduced form of vitamin K: Vitamin K is an enzymatic cofactor required for the post-translational modification of vitamin K-dependent (VKD) proteins. Although there are many VKD proteins, many of them are clinically relevant to patients with ESRD. They include central coagulation factors such as factors II, VII, IX, and X, as well as intercellular matrix proteins including matrix GLA-1 and osteocalcin. Under normal circumstances, vitamin K is reduced to vitamin K hydroquinone (KH2) by the enzyme NADPH oxidase. Only the reduced form of vitamin K can function as a cofactor for gamma glutamate carboxylase (GGCX), which catalyzes the carboxylation of vitamin K-dependent proteins. Warfarin blocks the production of vitamin K hydroquinone by acting as a reduction sink. Enzymatic carboxylation of glutamic acid residues results in further oxidation of vitamin KH2 to the 2-3 epoxide vitamin K ( Figure 2 ). The final step of the vitamin K cycle requires enzymatic oxidation of the vitamin K 2-3 epoxide back to its native structure. This step is catalyzed by vitamin K oxidoreductase (VKOR) and is an integral part of the vitamin K cycle, and is also blocked by the oxidative action of warfarin. The observation that warfarin blocks the production of vitamin K hydroxyquinone (KH2) and blocks the regeneration of vitamin K2 2-3 epoxide helps explain why calciphylaxis and other forms of malnutrition calcification occur more frequently in patients receiving warfarin therapy.

[0118] In one variation, administration or supplementation of the disclosed compounds and compositions reduces the risk of vascular and soft tissue calcification by increasing the formation of primary calcium binding protein particles (CPPs) composed of fetuin A and carboxylated matrix GLA-1 protein. Under normal physiological conditions, plasma calcium and phosphate concentrations are close to supersaturation, and are therefore expected to precipitate as crystalline hydroxyapatite in blood vessels and soft tissues. The observation that this process does not occur suggests that there are potent chemical and biological ways to block pathological calcification. Recent studies have shown that circulating calcium phosphate crystals are complexed with two calcification inhibitory proteins to form primary calcium binding protein particles (CPPs). These protein-mineral complexes are primarily composed of fetuin A; a protein derived from the liver that has been shown to prevent vascular calcification. The second protein, which is present at lower levels, is matrix Gla-1 protein, which is also used to prevent pathological calcification. Matrix Gla-1 is a vitamin K-dependent protein, and early work by Price et al. and others has shown that the formation of fetuin-matrix Gla-1 mineral nanoparticles (primary calcium binding protein CPPs) depends on the gamma carboxylation of matrix Gla-1. Preclinical studies have shown that the calcium-binding protein system serves as an alternative way to prevent pathological calcification when humoral defenses such as pyrophosphate, magnesium, and albumin are overwhelmed. The "absorption" of calcium phosphate crystals by primary CCPs occurs in a coordinated and time-dependent process.

[0119] The time for the primary CCP to reach 50% saturation (T 50 ) is an accurate and highly sensitive way to determine the ability of plasma to "sink" or "absorb" excess calcium phosphate crystals. 50 Patients with a short time showed a reduced ability to absorb calcium phosphate crystals, while T 50 Patients with longer durations are consistent with high ability. Recent clinical studies have confirmed that T 50 Tested and confirmed low T 50 Time is associated with increased myocardial infarction, heart failure, and all-cause mortality. Therefore, any clinical intervention that can increase the synthesis of circulating primary CCP will improve the ability to prevent pathological calcification. It is worth noting that since patients with CKD and ESRD show reduced levels of carboxylated matrix Gla-1 protein, and this process is essential for the formation of primary CPP. Therefore, supplementing or administering the disclosed compounds and compositions in CKD or ESRD patients will reduce the risk of pathological calcification and prevent the development of vascular and soft tissue calcification. Supplementing or administering the disclosed compounds or compositions can prevent or delay the development of soft tissue and vascular calcification in skin tissue by restoring the production of carboxylated matrix Gla-1 and GAS-6.

[0120] The regeneration of vitamin K involves two key enzymes: vitamin K 2-3 epoxide oxidoreductase (VKOR) and NAD(P)H:quinone oxidoreductase (NQO1). As shown in the figure (Figure), VKOR reduces 2-3 vitamin K epoxide to vitamin K quinone, while NADPH reduces vitamin K quinone to its hydroxyquinone form (KH2). Recent studies have shown that there are two different isoforms of VKOR that differ in both enzymatic properties and tissue distribution (VKORC-1 and VKORC1-like 1 [VKORC1-L1]). For example, Westhofen et al. have demonstrated that VKOCR-L1 has a 3-fold reduced affinity for 2-3 epoxide vitamin K compared to VKORC1. Subsequent studies supported the hypothesis that VKOR-L1 is a special isoform that prevents oxidative damage by regenerating vitamin K. When cultured HEK 293T cells were incubated with H2O2, expression of VKOR-L1 increased and evidence of membrane oxidative damage decreased. Clinical observations indicate that calciphylaxis and vitamin K-dependent vascular calcification are more common in the dermis, raising the question of whether the VKOR enzyme is differentially expressed in the skin. Casper et al. measured mRNA expression of key enzymes involved in vitamin K regeneration. In addition, expression of NADPH in the dermis was below detection. These observations suggest that any condition or procedure that impedes vitamin K remodeling (i.e., hemodialysis) would predispose the tissue to pathological calcification.

[0121] The oxidative properties of uremic plasma, as well as the oxidative effects of dialysis itself, lead to a “metabolic block” and accumulation of 2-3-epoxide vitamin K, as well as a reduction in intracellular levels of vitamin K2. The “downstream” effects of this block include the inability of key gamma carboxylate proteins to participate in the prevention of soft tissue and vascular calcification. This effect is exacerbated by the oxidative effects of hemodialysis, which may partially explain the propensity of ESRD patients to develop calciphylaxis and vascular calcification.

[0122] Relationship between Vitamin K and Circulating Vitamin K-Dependent Proteins in CKD-ESRD Patients: It is generally accepted that vitamin K levels may not be reduced in ESRD patients despite dietary deficiencies. For example, Holder et al. studied 172 stable dialysis patients and found that only 6% of patients exhibited clinically significant vitamin K deficiency. However, when the patients were examined for carboxylated osteocalcin levels, fully 60% had reduced levels. To confirm that this was a general effect of reduced vitamin K activity, the authors also measured PIVKA-II; another vitamin K-dependent protein. In fact, reduced levels of carboxylated prothrombin were found in 90% of both CKD and ESRD patients. In a similar study, Pilkey et al. measured vitamin K1 levels in 142 ESRD patients and found that most patients had adequate vitamin K stores, but 93% had non-carboxylated osteocalcin levels above 20% of total levels. There was no correlation between total vitamin K1 and circulating levels of non-carboxylated osteocalcin. This unexpected finding is consistent with the hypothesis that in uremic patients, total vitamin K levels can be normal, whereas the production of the reduced form is blocked by the oxidative properties of uremic disease.

[0123] In one variation, administration or supplementation or administration of the disclosed compounds and compositions will reverse hemodialysis-induced inhibition of vitamin K-dependent proteins by normalizing the functional reduced form of vitamin K. The observation that oxidative conditions may disrupt vitamin K circulation suggests that the oxidative load generated during hemodialysis may contribute to the high rates of vascular and soft tissue calcification observed within the ESRD population. Studies by Himmelfarb et al. and others have demonstrated that simple delivery of hemodialysis may result in oxidation of many tissue proteins. For example, hydroxy amino acid side chains are oxidized to carbonyl groups. In a study of CKD and ESRD patients, Himmelfarb et al. demonstrated the use of carbonyl side chain oxidation as a measure of overall oxidative burden. Himmelharb et al. showed that both CKD and ESRD patients exhibited a higher percentage of carbonyl proteins (15-fold) compared to normal controls (see Figure 5 ). In patients receiving dialysis, the percentage of carbonyl proteins was even higher, suggesting that dialysis not only reduces the oxidative burden but also appears to contribute to it. Figure 5As shown, patients with uremia were found to have up to 15-fold higher levels of carbonylated proteins. Thus, the oxidative load generated by hemodialysis delivery results in oxidation of functional vitamin K hydroquinone (KH2) to the non-functional native vitamin. Oxidation of KH2 by hemodialysis blocks its role as a cofactor for GGCX, resulting in reduced gamma carboxylation of downstream vitamin K-dependent proteins.

[0124] To confirm that uremia and hemodialysis can disrupt the vitamin K cycle, the ratio of vitamin K quinone to 2-3 epoxide vitamin K and vitamin K hydroxyquinone (KH2) can be measured in patients with normal renal function, CKD (stage IV & V) and ESRD patients. In order to determine whether the same hemodialysis process will further disrupt the vitamin K cycle, the level of oxidized vitamin K can be measured before hemodialysis, then in the middle of dialysis (2 hours) and 30 minutes after dialysis. Previous studies have examined the interaction between warfarin and vitamin K metabolism, and it has been shown that 2-3 epoxide vitamin K is easy to measure. Compared with controls, patients with CKD and ESRD will have higher levels of 2-3 epoxide vitamin K and lower levels of vitamin hydroquinone (KH2). In order to determine whether the loss of the reduced form of vitamin K (KH2) will lead to reduced carboxylation of vitamin K-dependent proteins, the levels of the following biomarkers in controls, CKD (stage IV and V) and ESRD (before and after hemodialysis) can be measured. Matrix GLA-1 protein; growth arrest-specific gene 6 (Gas-6) protein; PIVAK-II protein; osteocalcin; protein C; protein S; fetuin A; and osteoprotegerin (dialysis plasma level: 6.7 + 2.2 pmole / L). These studies were extended to include patients on stable 3x weekly hemodialysis. The levels of carboxylated vitamin K-dependent and non-carboxylated vitamin K-dependent proteins in pre-dialysis serum could be compared with levels obtained at 2 hours of dialysis and at the end of dialysis. The oxidative effects of dialysis itself would result in a decrease in the levels of carboxylated vitamin K2-dependent proteins.

[0125] In one variation, administration or supplementation of the disclosed compounds and compositions in ESRD patients with calcific uremic arteriole disease (calciphylaxis) will shorten wound healing time by preventing new vascular calcification and restoring blood flow: Skin biopsy: To confirm that supplementation with the disclosed compounds and compositions prevents the development of small vessel calcification and skin ischemia, patients with calciphylaxis confirmed by skin biopsy can be identified and randomized to treatment with menaquinone-7 or placebo. In another variation of the above procedure, treatment is with menaquinone-9 or placebo. Clinical endpoints may include the following: 1) time to withdrawal of wound vacuum therapy and 2) time to wound healing, defined as the time required for a 50% reduction in the total surface area of ​​all calciphylaxis wounds.

[0126] In another variation, administration or supplementation of the disclosed compounds and compositions significantly increases the bioavailability of the following compounds and their respective mixtures when compared to administration or supplementation: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9, and menaquinone-10 / naphthol-10. In one variation, the bioavailability is increased by at least 5%, 10%, 15%, 20%, 30%, 40%, 50% or more when compared to administration or supplementation with menaquinone-7, menaquinone-8, menaquinone-9, and menaquinone-10, respectively.

[0127] In another variation, administration or supplementation of the disclosed compounds and compositions significantly increases the serum half-life of the following compounds and their respective mixtures when compared to administration or supplementation: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9, and menaquinone-10 / naphthol-10. In one variation, the serum half-life is increased by at least 5%, 10%, 15%, 20%, 30%, 40%, 50% or more when compared to administration or supplementation with menaquinone-7, menaquinone-8, menaquinone-9, and menaquinone-10, respectively.

[0128] Histopathological endpoints: Comparison of diagnostic skin biopsy after 12 weeks of menadione-7 therapy with protocol repeat skin biopsy. Changes in interstitial calcium deposition levels are defined as changes in Von Kossa staining, which can be quantified by digital image color analysis. Skin biopsies can be used to validate biomarkers at the tissue level. This process can confirm the preventive properties of the disclosed menadione derivatives against early vascular calcification. Validation of these biomarkers in tissue will also enable clinicians to utilize biomarkers as a way to track clinical responsiveness. Microvascular calcification precedes the development of CUA lesions. Calcification levels can be quantified by Von Kossa calcium staining in peripheral tissues and normalized to calcium content per unit area. Von Kossa can be used as a way to confirm the preventive properties of the disclosed menadione derivatives against the development of vascular calcification. In one variation, supplementation of the disclosed compounds and compositions in ESRD patients with calcific uremic arteriopathy (calciphylaxis; CUA) will shorten wound healing time by normalizing carboxyprotein C levels in the dermis and preventing primary thrombosis of cutaneous vessels. Thus, in one variation, supplementation or administration of the disclosed compounds or compositions in diabetic patients will prevent the development of vascular dementia by preventing the development of calcification and small vessel lesions. In another embodiment, a fortified food or beverage formulation is provided, comprising adding a composition comprising a compound or a mixture thereof comprising any of the above compounds to food or beverage.

[0129] Also included in the above embodiments, aspects and modifications are amino acid salts (such as arginine salts, etc.), gluconates and galacturonates. Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms, and are intended to fall within the scope of the present invention. Also provided is a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a therapeutically effective amount of at least one compound of the present invention.

[0130] The pharmaceutical composition of the compound of the present invention or its derivative can be formulated into a solution or lyophilized powder for parenteral administration. The powder can be reconstituted by adding a suitable diluent or other pharmaceutically acceptable carrier before use. Liquid preparations are generally buffered isotonic aqueous solutions. Examples of suitable diluents are isotonic physiological saline solutions, 5% glucose aqueous solutions, or buffered sodium acetate or ammonium acetate solutions. Such preparations are particularly suitable for parenteral administration, but can also be used for oral administration. Excipients such as polyvinyl pyrrolidone, gelatin, hydroxycellulose, gum arabic, polyethylene glycol, mannitol, sodium chloride or sodium citrate can also be added. Alternatively, these compounds can be encapsulated, tableted or made into emulsions or syrups for oral administration. Pharmaceutically acceptable solid or liquid carriers can be added to enhance or stabilize the composition, or to promote the preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, glycerol, saline, alcohol or water. Solid carriers include starch, lactose, calcium sulfate, dihydrate, clay, magnesium stearate or stearic acid, talc, pectin, gum arabic, agar or gelatin. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies, but may be between about 20 mg and about 1 g per dosage unit. The pharmaceutical preparation is prepared according to conventional pharmaceutical techniques, involving grinding, mixing, granulating and compressing for tablet form, if necessary; or grinding, mixing and filling for hard gelatin capsule form. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or aqueous or non-aqueous suspension. Such liquid preparations can be directly administered po or filled into soft gelatin capsules. Suitable formulations for each of these methods of administration can be found, for example, in Remington: The Science and Practice of Pharmacy, A. Gennaro, ed., 20th ed., Lippincott, Williams & Wilkins, Philadelphia, Pa.

[0131] The disclosed compounds and compositions may include a solubility enhancer or solubilizer selected from the group consisting of oleic acid, EL (polyoxyethylene castor oil or Cremophor EL), Vitamin E TPGS (D-α-tocopheryl polyethylene glycol-1000 succinate), Savie (a polysarcosine-derived emulsifier), CC (monolinoleylglycerol), 44 / 14 (Lauroyl polyoxyethylene-32 glyceryl), 812N (esters of caprylic fatty acids and glycerol derived from saturated coconut and palm kernel oils), Oleique (polyglyceryl-6 dioleate), Lauroglycol TM 90(propylene glycol monolaurate (type II), (Caprylocaproyl Polyoxyethylene-8 Glyceryl), (SBE-β-cyclodextrin), Encapsin TM HPB (Hydroxypropyl β-cyclodextrin), Peceol TM (Glycerol / Glycerol Monooleate (Type 40)), Sodium Deoxycholate, Deoxycholic Acid, M2125CS (linoleoyl polyoxyl-6 glyceride) and medium chain mono- and di-glycerides.

[0132] In one variation, provided are compounds disclosed herein, or pharmaceutically acceptable salts thereof, optionally in the form of a single stereoisomer or a mixture of stereoisomers thereof; and compositions comprising the same.

[0133] In addition to the exemplary embodiments, aspects, and variations described above, further embodiments, aspects, and variations will become apparent by reference to the drawings and by examination of the following descriptions. DETAILED DESCRIPTION

[0134] definition:

[0135] Unless otherwise specifically stated herein, the definitions of terms used are standard definitions used in the fields of organic synthesis and pharmaceutical sciences.Exemplary embodiments, aspects, and variations are shown in the accompanying drawings, and it is intended that the embodiments, aspects, and variations disclosed herein and the drawings be considered illustrative rather than limiting.

[0136] "Pharmaceutically acceptable salt" means a salt composition generally recognized to have the desired pharmacological activity, which is considered safe, non-toxic and acceptable for use in veterinary and human medicine. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.; or with organic acids such as acetic acid, propionic acid, caproic acid, malonic acid, succinic acid, malic acid, citric acid, gluconic acid, salicylic acid, etc.

[0137] "Therapeutically effective amount" means the amount of a compound or drug that elicits any one of the biological effects listed in the specification. BRIEF DESCRIPTION OF THE DRAWINGS

[0138] Figure 1 is a representation of the chromatogram of menaquinone-7 and its positional isomers shown in a ratio of 3:1, by 1 Determined by H NMR.

[0139] Figure 2 Schematic diagram showing uremia- and dialysis-induced KH2 oxidation, the functional carboxylation of vitamin K-dependent proteins.

[0140] Figure 3 Graphs showing VKORC1 in arbitrary units and in specific tissues.

[0141] Figure 4 is a graph showing NADPH in arbitrary units and in specific tissues.

[0142] Figure 5 is a graph showing that CKD and ESRD patients exhibit a higher percentage of carbonyl proteins compared to normal controls. Experimental:

[0143] The following procedures can be used to prepare the compounds of the present invention. Starting materials and reagents used to prepare these compounds are available from commercial suppliers such as Aldrich Chemical Company (Milwaukee, Wis.), Bachem (Torrance, Calif.), Sigma (St. Louis, Mo.), or are prepared by methods well known to those of ordinary skill in the art following the procedures described in such references as Fieser and Fieser's Reagents for Organic Synthesis, Vols. 1-17, John Wiley and Sons, New York, NY, 1991; Rodd's Chemistry of Carbon Compounds, Vols. 1-5 and Supplements, Elsevier Science Publishers, 1989; Organic Reactions, Vols. 1-17, John Wiley and Sons, New York, NY, 1991; Rodd's Chemistry of Carbon Compounds, Vols. 1-5 and Supplements, Elsevier Science Publishers, 1989; Reactions, vols. 1-40, John Wiley & Sons, New York, NY, 1991; March J.: Advanced Organic Chemistry, 4th ed., John Wiley & Sons, New York, NY; and Larock: Comprehensive Organic Transformations, VCH Press, New York, 1989.

[0144] In some cases, protecting groups may be introduced and eventually removed. Suitable protecting groups for amino, hydroxyl and carboxyl groups are described in Greene et al., Protective Groups in Organic Synthesis, 2nd ed., John Wiley & Sons, New York, 1991. Standard organic chemistry reactions can be achieved using a variety of different reagents, for example, as described in Larock: Comprehensive Organic Transformations, VCH Press, New York, 1989.

[0145] Preparation of naphthol derivatives:

[0146] General reaction scheme:

[0147]

[0148] m and m 1 = an integer from 3 to 15;

[0149] n = 7, 8, 9 or 10;

[0150] R 1 =R 1 is H or -CH3;

[0151] s = 1 or 2;

[0152] X = halide (Cl, Br, I), -OC(O)C1-C6 alkyl, -OC(O)(CH2) q -OC(O)C1-C6 alkyl and -OZ-(C=Y)(CH2) s -(OCH2CH2)-OR 1

[0153] Y and Y 1 =O, NH or S; and

[0154] Z and Z 1 =-C(O)CH2O-, -C(O)CH2NH- or absent.

[0155] The preparation of naphthol monoester and diester derivatives can be carried out using standard methods known in the art and as described herein. For example, as shown in the general reaction scheme, menaquinone (such as MK-7 or MK-9) can be pegylated with an acid halide in an organic solvent (such as DCM, hexane, toluene and THF) and in the presence of an organic base or an inorganic base (such as Et3N, pyridine, DMAP, NaOAc, NaHCO3 and zinc). A (X = chloride, bromide or iodide). Alternatively, acylation of the quinone can be carried out with symmetrical or unsymmetrical pegylated anhydrides and catalytic acids such as H2SO4, HCl and phosphoric acid.

[0156] For example, menaquinones (such as MK-7 or MK-9) can be pegylated with anhydrides in organic solvents (such as DCM, hexane, toluene, and THF) and in the presence of organic or inorganic bases or metals (such as Et3N, pyridine, DMAP, NaOAc, NaHCO3, and zinc). A (X=-OC(O)R 2 , where R 2 Under certain conditions, using stoichiometric amounts of acylating agent, the manner in which the acylating agent is added to the menadione, and the reaction conditions, selective acylation can be achieved to provide monoacylated esters at the 1-position or 4-position of the menadione.

[0157] Alternatively, the preparation of naphthol monoester and diester derivatives can be carried out using HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxide hexafluorophosphate, azabenzotriazole tetramethyluronium hexafluorophosphate) as a coupling agent, wherein the corresponding PEGylated carboxylic acid A (X = -OH) is treated with HATU to form an -OAt active ester, which is then reacted with the corresponding naphthol formed from menadione. See, e.g., Carpino La et al., The uronium / guanidinium peptide coupling reagents: Finally the true uranium salts. Angewandte Chemie International Edition, 2002, 41(3): 441-445; Carpino LA et al., Comparison of the Effects of 5-and 6-HOAt on Model Peptide Coupling Reactions Relative to the Cases for the 4-and7-Isomers. Organic letters, 2000, 2(15): 2253-2256.

[0158] Selective hydrolysis of diesters:

[0159]

[0160] In addition, the selective formation of monoester naphthol at the 1-position or 4-position can be carried out by initially preparing a diester and then selectively hydrolyzing one of the ester groups to form the desired monoester derivative. For example, it has been shown that under certain controlled conditions, the naphthol ester formed at the 4-position of the quinone (a ketone adjacent to the isoprenoid side chain) is more easily hydrolyzed than the naphthol ester group at the 1-position. Standard hydrolysis conditions include aqueous NaOH in the presence of an organic solvent (such as THF or diethyl ether), or THF and / or methanol or ethanol containing NaOMe. Alternatively, the hydrolysis can be carried out in a dilute acid aqueous solution, such as HCl or H2SO4, optionally in an organic solvent or a mixture of organic solvents at high temperature, such as THF or ether, such as about 30°C to 50°C. When a mixture of monoesters and diesters is formed by a hydrolysis reaction, i.e., an incomplete selective hydrolysis, the desired monoester can be easily separated from the diester by column chromatography or preparative chromatography.

[0161] Acylation and selective hydrolysis reactions carried out under various standard conditions known in the art provide the desired mono-acylated or di-acylated derivatives having the desired 1 H-NMR spectrum name.

[0162] General reaction scheme:

[0163]

[0164] Experimental Procedure:

[0165] Step 1: Preparation of 4-(acetoxy)-2-methylnaphth-1-yl acetate (2):

[0166]

[0167] To a stirred solution of Na2S2O4 (42.4 g, 0.24 mol) in water (160 mL, 8 V) was added ethyl acetate (160 mL, 8 V) containing 2-methyl-1,4-dihydronaphthalene-1,4-dione 1 (20 g, 0.116 mol) at room temperature (25-30 ° C). The reaction mixture was stirred at room temperature for 2 hours. The completion of the reaction was observed by the color change from a dark brown solution to a light brown solution. The reaction mixture was transferred to a separatory funnel, and the separated organic layer was dried over sodium sulfate and concentrated. The obtained crude product was placed in acetic anhydride (66 mL, 0.69 mol), cooled to 0 ° C, and concentrated H2SO4 (4 mL, 0.2 V) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC to be complete, R f =0.4 (20% ethyl acetate / hexane).

[0168] The reaction mixture was slowly added to water (500 mL) at ambient temperature (25-30° C.) and stirred at the same temperature (25-30° C.) for 10 minutes. The obtained solid was filtered and dried to give compound 2 (26.0 g, 87%) as a brown solid. f =0.4 (20% ethyl acetate / hexane). 1 H NMR (400 MHz, CDCl3) δ (ppm): 7.86-7.80 (m, 1H), 7.80-7.72 (m, 1H), 7.57-7.44 (m, 2H), 7.15 (s, 1H), 2.47 (d, J = 11.0 Hz, 6H), 2.33 (s, 3H). LC-MS m / z (M+1): 259.8 (calculated molecular weight -258.26).

[0169] Step 2: Preparation of 4-hydroxy-2-methylnaphthalen-1-yl acetate (3):

[0170]

[0171] Sodium hydroxide (3.8 g, 0.096 mol) and Na2S2O4 (5.0 g, 0.029 mol) aqueous solution (50 mL, 2 V) were added to a stirred solution of compound 2 (25 g, 0.096 mol) in ethanol (500 mL, 20 V) at room temperature (25-30 ° C). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored to completion by TLC. The reaction mixture was neutralized with 1.5 N HCl (pH-3 to 4), the resulting reaction mixture was filtered, and the filtrate was concentrated (to remove ethanol). The crude product was diluted with ethyl acetate (500 mL), washed with water (2X 200 mL) and brine solution (200 mL). The separated organic layer was dried over sodium sulfate and concentrated. The crude product was washed with n-hexane (3X 100 mL) to obtain compound 3 (18.2 g, 87%) as a brown solid. f =0.3 (20% ethyl acetate / hexane). 1 H NMR (400 MHz, CDCl3) δ (ppm): 8.03 (dt, J = 8.3, 1.1 Hz, 1H), 7.70-7.61 (m, 1H), 7.49 (ddd, J = 8.4, 6.8, 1.4 Hz, 1H), 7.41 (ddd, J = 8.3, 6.8, 1.3 Hz, 1H), 6.48 (s, 1H), 5.54 (d, J = 1.5 Hz, 1H), 2.49 (s, 3H), 2.21 (s, 3H). LC-MS m / z (M+1): 217.6 (calculated molecular weight -216.23).

[0172] Synthesis of MK-7-monoacetate:

[0173] Step 3: Preparation of 3-[(2E,6E,10E,14E,18E,22E)-3,7,11,15,19,23,27-heptamethyloctacosane-2,6,10,14,18,22,26-heptaen-1-yl]-4-hydroxy-2-methylnaphthalen-1-yl acetate (4):

[0174]

[0175] Heptyl isopentenol (16.0 g, 0.032 mol) and benzenesulfonic acid (0.72 g, 0.0046 mol) were added to a stirred solution of compound 3 (10.0 g, 0.046 mol) in toluene (100 mL, 10 V) at room temperature (25-30 ° C). The reaction mixture was stirred at room temperature for 16-24 hours. The reaction was monitored to completion by TLC. The reaction mixture was diluted with ethyl acetate (500 mL), washed with water (2X 250 mL) and saline solution (200 mL). The separated organic layer was dried over sodium sulfate and concentrated. The crude product obtained was purified by column chromatography (2-3% ethyl acetate / hexane), and crystallized from a combination of acetone and n-heptane (1: 4 V), to obtain compound 4 (4.1 g, 13%) as an off-white solid, R f =0.5 (15% ethyl acetate / heptane). 1 H NMR (400MHz, CDCl3) δ (ppm): 8.13-8.10 (m, 1H), 7.64-7.62 (m, 1H), 7.46-7.39 (m, 2H), 5.70 (s, 1H), 5.27-5.24 (m, 1H), 5.11 (tp, J=6.8, 4.4, 2.9H z,6H),3.53(d,J=6.8Hz,2H),2.47(s,3H),2.26(s,3H),2.17-1.93(m,24 H), 1.88 (d, J = 1.3Hz, 3H), 1.68 (d, J = 1.6Hz, 3H), 1.59 (d, J = 9.7Hz, 18H). LC-MS m / z (M+1): 693.39 (calculated monoisotopic mass -692.51).

[0176] Synthesis of MK-7-PEG2-monoacetate:

[0177] Step 4: Preparation of 4-(acetoxy)-2-[(2E,6E,10E,14E,18E,22E)-3,7,11,15,19,23,27-heptamethyloctacosane-2,6,10,14,18,22,26-heptaen-1-yl]-3-methylnaphthalen-1-yl 2-[2-(2-methoxyethoxy)ethoxy]acetate:

[0178]

[0179] At 0-5 ℃, N, N'-dicyclohexylcarbodiimide (0.534g, 0.0025mol) and 4-dimethylaminopyridine (17mg, 0.00014mol) were added to a solution of compound 5 (0.41g, 0.00230mol) in DCM (10mL, 10V), and the reaction mixture was stirred at the same temperature (0-5 ℃) for 10 minutes. DCM (10mL) containing compound 4 (1.0g, 0.0014mol) was added to the reaction mixture at 0-5 ℃, and slowly stirred at ambient temperature for 2 hours at 25-30 ℃. The reaction was monitored to completion by TLC. The reaction mixture was diluted with dichloromethane (100mL), washed with water (2X50mL) and saline solution (100mL). The separated organic layer was dried over sodium sulfate and concentrated. The obtained crude product was purified by column chromatography (20% ethyl acetate / hexane) to give compound MK-7-PEG2-monoacetate (738 mg, 60%) as a light yellow viscous liquid. f =0.4 (20% ethyl acetate / hexane). 1 H NMR (400 MHz, CDCl3) δ (ppm): 7.74-7.60 (m, 2H), 7.51-7.41 (m, 2H), 5.15-5.01 (m, 7H), 4.62 (s, 2H), 3.93-3.86 (m, 2H), 3.80-3.65 (m, 4H), 3.61-3.54 (m, 2H), 3.39 (s, 5H), 2.48 (s, 3H), 2.24 (s, 3H), 2.10-1.90 (m, 24H), 1.72 (dd, J = 35.3, 1.3 Hz, 6H), 1.57 (d, J = 1.7 Hz, 18H). LC-MS m / z (M+1): 853.39 (calculated monoisotopic mass -852.59). HPLC purity: 99.58%.

[0180] Synthesis of MK-7-PEG4-monoacetate:

[0181] Step 4: Preparation of 4-(acetoxy)-2-[(2E,6E,10E,14E,18E,22E)-3,7,11,15,19,23,27-heptamethylcarbocosanoic acid-2,6,10,14,18,22,26-heptaen-1-yl]-3-methylnaphthalen-1-yl 2,5,8,11,14-pentaoxahexadecan-16-oic acid ester:

[0182]

[0183] 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (0.497g, 0.0025mol) and 4-dimethylaminopyridine (17mg, 0.00014mol) were added to a solution of compound 5 (0.614g, 0.00230mol) in DCM (10mL, 10V) at 0-5°C, and the reaction mixture was stirred at the same temperature (0-5°C) for 10 minutes. DCM (10mL) containing compound 4 (1.0g, 0.0014mol) was added to the reaction mixture at 0-5°C, and slowly stirred for 2 hours at ambient temperature 25-30°C. The reaction was monitored by TLC to complete. The reaction mixture was diluted with dichloromethane (100mL), washed with water (2X 50mL) and saline solution (100mL). The separated organic layer was dried over sodium sulfate and concentrated. The obtained crude product was purified by column chromatography (40% ethyl acetate / hexane) to give compound MK-7-PEG4-monoacetate (842 mg, 62%) as a light yellow viscous liquid. f =0.4 (40% ethyl acetate / hexane). 1 H NMR(400MHz, CDCl3)δ(ppm):7.76-7.59(m,2H),7.55-7.41(m,2H),5.19-4.99(m,7H),4.62 (s,2H),3.88(dd,J=5.8,3.5Hz,2H),3.75(dd,J=5.7,3.5Hz,2H),3.66(ddt,J=15.1,9.2,3 .6 Hz, 10H), 3.53 (dd, J = 5.8, 3.6 Hz, 2H), 3.36 (s, 5H), 2.48 (s, 3H), 2.24 (s, 3H), 2.01 (dtd, J = 31.6, 8.5, 7.9, 4.8 Hz, 24H), 1.76 (d, J = 1.3 Hz, 3H), 1.68 (d, J = 1.5 Hz, 3H), 1.57 (s, 18H). LC-MS m / z (M+1): 941.31 (calculated monoisotopic mass -940.64). HPLC purity: 99.62%.

[0184] Synthesis of MK-9-monoacetate:

[0185] General reaction scheme:

[0186]

[0187] Step 3: Preparation of 4-hydroxy-2-methyl-3-[(2E,6E,10E,14E,18E,22E,26E,30E)-3,7,11,15,19,23,27,31,35-nonamethylhexatriacontane-2,6,10,14,18,22,26,30,34-nonaen-1-yl]naphthalen-1-yl acetate (4):

[0188]

[0189] Solanesol (20.45 g, 0.032 mol) and benzenesulfonic acid (0.72 g, 0.0046 mol) were added to a stirred solution of compound 3 (10.0 g, 0.046 mol) in toluene (100 mL, 10 V) at room temperature (25-30 ° C). The reaction mixture was stirred at room temperature for 16-24 hours. The reaction was monitored to completion by TLC (15% ethyl acetate / hexane). The reaction mixture was diluted with ethyl acetate (500 mL), washed with water (2X 250 mL) and brine solution (200 mL). The separated organic layer was dried over sodium sulfate and concentrated. The crude product obtained was purified by column chromatography (2-3% ethyl acetate / hexane) and crystallized from a combination of acetone and n-heptane to obtain compound 4 (4.1 g, 13%) as an off-white solid, R f =0.5 (15% ethyl acetate / heptane). 1 H NMR (400MHz, CDCl3) δ (ppm): 8.13-8.10 (m, 1H), 7.64-7.62 (m, 1H), 7.46-7.26 (m, 2H), 5.70 (S, 1H), 5.28-5.24 (m, 1H), 5.11 (tp, J=6.8, 4.4, 2.9H z,9H),3.53(d,J=6.8Hz,2H),2.47(s,3H),2.26(s,3H),2.17-1.93(m,32 H), 1.88 (d, J = 1.3Hz, 3H), 1.68 (d, J = 1.6Hz, 3H), 1.59 (d, J = 9.7Hz, 24H). LC-MS m / z (M+1): 829.36 (calculated monoisotopic mass -828.64).

[0190] Synthesis of MK-9-PEG2-monoacetate:

[0191] Step 4: 4-(Acetoxy)-3-methyl

[0192] Preparation of 2-[(2E,6E,10E,14E,18E,22E,26E,30E)-3,7,11,15,19,23,27,31,35-nonamethylhexatriacontane-2,6,10,14,18,22,26,30,34-nonaen-1-yl]naphthalen-1-yl 2-[2-(2-methoxyethoxy)ethoxy]acetate:

[0193]

[0194] N, N'-dicyclohexylcarbodiimide (0.447g, 0.0021mol) and 4-dimethylaminopyridine (14mg, 0.00012mol) were added to a solution of compound 5 (0.513g, 0.00192mol) in DCM (10mL, 10V) at 0-5°C, and the reaction mixture was stirred at the same temperature (0-5°C) for 10 minutes. DCM (10mL) containing compound 4 (1.0g, 0.0012mol) was added to the reaction mixture at 0-5°C. The reaction mixture was slowly stirred at ambient temperature for 2 hours at 25-30°C. The reaction was monitored by TLC to complete. The reaction solution was diluted with dichloromethane (100mL), washed with water (2X 50mL) and saline solution (100mL). The separated organic layer was dried over sodium sulfate and concentrated. The crude product obtained was purified by column chromatography (20% ethyl acetate / hexane) to give compound MK-9-PEG2-monoacetate (738 mg, 60%) as an off-white solid, Rf=0.4 (20% ethyl acetate / hexane). 1 H NMR (400MHz, CDCl3) δ (ppm): 7.71-7.63 (m, 2H), 7.47-7.44 (m, 2H), 5.12-5.00 (m, 9H), 4.62 (s, 2H), 3.90-3.86 (m, 2H), 3.80-3.65 (m, 4H), 3.6 1-3.54(m,2H),3.39(s,5H),2.48(s,3H),2.24(s,3H),2.10-1.91(m,3 2H), 1.77 (d, J = 1.4Hz, 3H), 1.68 (t, J = 1.4Hz, 3H), 1.60-1.53 ​​(m, 24H). LC-MS m / z (M+1): 989.75 (calculated monoisotopic mass -988.71). HPLC purity: 99.18%.

[0195] Synthesis of MK-9-PEG4-monoacetate:

[0196] Step 4: 4-(Acetoxy)-3-methyl

[0197] Preparation of 2-[(2E,6E,10E,14E,18E,22E,26E,30E)-3,7,11,15,19,23,27,31,35-nonamethylhexatriacontane-2,6,10,14,18,22,26,30,34-nonaen-1-yl]naphthalene-1-yl 2,5,8,11,14-pentaoxahexadecan-16-acid ester:

[0198]

[0199] 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (0.416g, 0.00217mol) and 4-dimethylaminopyridine (17mg, 0.00014mol) were added to a solution of compound 5 (0.513g, 0.00192mol) in DCM (2.0L, 400V) at 0-5°C, and the reaction mixture was stirred at the same temperature (0-5°C) for 10 minutes. DCM (10mL) containing compound 4 (1.0g, 0.0012mol) was added to the above reaction mixture at 0-5°C. The reaction mixture was slowly stirred at ambient temperature for 2 hours at 25-30°C. The reaction mixture was monitored to completion by TLC. The reaction mixture was diluted with dichloromethane (100mL), washed with water (2X 50mL) and saline solution (100mL). The separated organic layer was dried over sodium sulfate and concentrated. The obtained crude product was purified by column chromatography (40% ethyl acetate / hexane) to give compound MK-9-PEG4-monoacetate (779 mg, 60%) as a light yellow viscous liquid, Rf=0.4 (40% ethyl acetate / hexane). 1 H NMR (400MHz, CDCl3) δ (ppm): 7.71-7.63 (m, 2H), δ 7.47-7.45 (m, 2H), 5.09 (dt, J = 20. 5,7.7Hz,9H),4.61(s,2H),3.88(dd,J=5.7,3.6Hz,2H),3.74(dd,J=5.7,3.6Hz,2H), 3.70-3.54 (m, 10H), 3.52 (dd, J = 5.9, 3.4 Hz, 2H), 3.37 (s, 5H), 2.48 (s, 3H), 2.24 (s, 3H), 2.02 (dt, J = 33.0, 7.5 Hz, 32H), 1.77 (s, 3H), 1.68 (d, J = 1.5 Hz, 3H), 1.60 (s, 24H). LC-MS m / z (M+1): 1077.75 (calculated monoisotopic mass -1076.76). HPLC purity: 98.75%

[0200] Synthesis of MK-9:

[0201] Step 1: Preparation of 2-methyl-3-[(2E,6E,10E,14E,18E,22E,26E,30E)-3,7,11,15,19,23,27,31,35-nonamethylhexatriacontane-2,6,10,14,18,22,26,30,34-nonaen-1-yl]-1,4-dihydronaphthalene-1,4-dione):

[0202]

[0203] Cerium ammonium nitrate (1.9 g, 0.0036 mol) was added to a stirred solution of compound 1 (1.0 g, 0.0012 mol) in a mixture of acetonitrile: ether: water (1:1:1) (12 mL, 12 V) at room temperature (25-30 ° C). The reaction mixture was stirred at room temperature for 1-2 hours. The reaction was monitored to completion by TLC. The reaction mixture was diluted with ethyl acetate (100 mL), washed with water (2X 50 mL) and saline solution (100 mL). The separated organic layer was dried over sodium sulfate and concentrated. The crude product obtained was purified by column chromatography (2-3% ethyl acetate / hexane) and crystallized from a combination of acetone and n-heptane (1:4 V) to obtain compound MK9 (580 mg, 51%) as a yellow solid, R f =0.7 (15% ethyl acetate / heptane). 1 H NMR (400 MHz, CDCl3) δ (ppm): 8.08 (ddd, J = 6.2, 3.1, 1.8 Hz, 2H), 7.71-7.66 (m, 2H), 5.20-4.99 (m, 9H), 3.37 (d, J = 7.0 Hz, 2H), 2.19 (s, 3H), 2.12-1.89 (m, 32H), 1.68 (d, J = 1.4 Hz, 3H), 1.66-1.51 (m, 24H). LC-MS m / z (M+1): 785.27 (calculated monoisotopic mass -784.61). HPLC purity: 98.0%.

[0204] The following representative compounds can be prepared using the above process:

[0205]

[0206]

[0207] Menaquinone-7 (MK-7) and compounds Ia.1 , m=9, n=7: Comparative single-dose oral pharmacokinetics study in male Sprague Dawley Rats:

[0208]

[0209] About 5 mg of menadione-7 was weighed and transferred to a graduated tube. Then 10 mL of sunflower oil was added and thoroughly vortexed and mixed until the test substance was completely dissolved. The preparation was prepared in a dark room under monochromatic light. The final strength of the preparation was 500 μg / mL. The preparation was freshly prepared before application.

[0210] PK study of oral menaquinone-7 in male SD rats: Clinical observations:

[0211]

[0212] Results: PK study of oral menaquinone-7 in male SD rats - Plasma concentrations of menaquinone-7:

[0213]

[0214]

[0215] Results: PK study of oral menaquinone-7 in male SD rats - Plasma PK parameters of menaquinone-7:

[0216]

[0217]

[0218] The compound is equivalent to 1000 μg menaquinone-7 per kg body weight. Ia.1 , m=9, n=7 Single-dose oral pharmacokinetic study in male Sprague Dawley rats:

[0219] Oral administration of compounds in male SD rats Ia.1 , m=9, n=7 PK study:

[0220]

[0221] Preparation: Oral administration of compounds in male SD rats Ia.1 , m=9, n=7 PK study:

[0222] Weigh approximately 6 mg of compound Ia.1 , m=9, n=7 and transferred to a graduated tube. Then add 9.836mL sunflower oil and vortex mix thoroughly until the test substance is completely dissolved. The preparation is prepared in a dark room under monochromatic light. The final concentration of the preparation is 610μg / mL (equivalent to 500μg / mL of MK-7). The preparation is freshly prepared before application.

[0223] Oral administration of compounds in male SD rats Ia.1 PK study of m=9, n=7: Clinical observation:

[0224]

[0225]

[0226] Results: Male SD rats were treated with oral administration of the compound Ia.1 PK study of compound Ia.1 , m=9, n=7 plasma concentration:

[0227]

[0228] Oral administration of compounds in male SD rats Ia.1 PK study with m=9 and n=7 - Plasma concentration of menaquinone-7:

[0229]

[0230]

[0231] Results: Male SD rats were treated with oral administration of the compound Ia.1 PK study with m=9 and n=7: Plasma PK parameters of MK-7:

[0232]

[0233] Results: Comparative Data - Plasma Concentrations of Menaquinone-7:

[0234]

[0235]

[0236] Comparative Data - Plasma PK Parameters of Menaquinone-7:

[0237]

[0238] Based on the above results, the maximum plasma concentration of menaquinone-7 in rats after oral administration of 1 mg / kg body weight using sunflower oil as a formulation vehicle was 19 ng / mL. Ia.1 , m=9, n=7. When given at comparable doses, the maximum plasma concentration observed was about 40% higher. Depending on the compound tested, the maximum plasma concentration was about 40% to 60% higher.

[0239] After administration of menaquinone-7, the observed plasma exposure of menaquinone-7 was 279.43 ± 74.91, while in the case of compound Ia.1, m = 9, n = 7, the plasma exposure of menaquinone-7 was about 45% higher. Compared with a similar dose of menaquinone-7 administered orally at 1000 μg / kg body weight alone, it was found that oral administration of the compound Ia.1 , m = 9, n = 7, the plasma exposure of menaquinone-7 was higher. No compound was observed Ia.1 , concentration of m=9, n=7.

[0240] Administration of the compound in subjects at risk for calciphylaxis:

[0241] This example describes the administration of the compounds of the present application to subjects who are at risk for developing calciphylaxis but have not yet developed the characteristic skin lesions of calciphylaxis. Risk factors to consider include, but are not limited to, diabetes, obesity, hemodialysis, and prior treatment with warfarin (Nigwekar et al. (2016) "A Nationally Representative Study of Calcific Uremic Arteriolopathy Risk Factors", Journal of the American Society of Nephrology (J. Am. Soc. Nephrol.) 27 (11): 3421-9). Administration of these compounds can protect subjects from skin lesions and change certain biomarker levels, indicating that the development of calciphylaxis can be prevented.

[0242] Subjects at risk for calciphylaxis receive 5 mg, 10 mg, 20 mg, 30 mg, 50 mg or 100 mg of the selected compound of the present application orally once daily for at least 2 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 6 months, 1 year or indefinitely. The dosage form is a 5 mg, 10 mg, 20 mg, 50 mg or 100 mg soft gel capsule. Two 50 mg capsules are administered once daily to the 100 mg dose cohort. All subjects with elevated risk factors for calciphylaxis will develop characteristic skin lesions of calciphylaxis. The purpose of active (before clinical diagnosis of calciphylaxis) treatment with the compounds of the present application is to prevent the appearance of lesions. A decrease in the frequency or elimination of lesions is considered a successful treatment.

[0243] Several biomarkers can be assessed to determine the efficacy of the compound to be administered at three dose levels. Exemplary biomarkers include PIVKA-II; uncarboxylated and total matrix Gla protein (MGP); uncarboxylated, carboxylated and total osteocalcin; uncarboxylated, carboxylated and total protein C, osteoprotegerin, fetuin A and hs-CRP. Blood samples are obtained according to the following schedule to measure biomarkers. Blood sampling can be performed weekly or monthly during treatment. It is envisioned that administration of the disclosed compounds will result in (i) a reduction in PIVKA-II, which indicates a delay in the progression of calciphylaxis, prevention or reversal of calciphylaxis, (ii) a reduction in uncarboxylated MGP, uncarboxylated osteocalcin and / or uncarboxylated protein C, which indicates a delay in the progression of calciphylaxis, prevention or reversal of calciphylaxis. Further, pulse wave velocity (PWV) can be measured to assess arterial compliance. An improvement in vascular compliance will indicate a delay in the progression of calciphylaxis, prevention or reversal of calciphylaxis.

[0244] Administration of compounds disclosed herein in subjects diagnosed with calciphylaxis:

[0245] This example describes the administration of a disclosed compound to a subject diagnosed with calciphylaxis. Typical symptoms include the appearance of characteristic painful skin lesions (Nigwekar et al. (2015) Calciphylaxis: Risk Factors, Diagnosis, and Treatment. Am. J. Kidney Dis. 66: 133-46). A definitive diagnosis of calciphylaxis can be made by skin biopsy. Additional circumstances need to be considered for a correct diagnosis.

[0246] Subjects diagnosed with calciphylaxis receive 5 mg, 10 mg, 25 mg, 50 mg, or 100 mg of a disclosed compound orally once daily for at least 2 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 6 months, 1 year, or indefinitely. The dosage form is a 5 mg, 10 mg, 25 mg, 50 mg, or 100 mg soft gel capsule. Two 50 mg capsules are administered once daily to the 100 mg dose cohort.

[0247] Prevention or reduction of lesion size and frequency is considered a sign of successful treatment. Administration of the compounds disclosed above will result in prevention or reduction of lesion size and frequency. In addition, since calciphylaxis carries a considerable risk of death, an increase in the overall survival time of the diagnosed subject will be a sign of successful treatment. In addition, administration of the compounds disclosed above will result in an increase in the overall survival time of the diagnosed subject.

[0248] Administration of the disclosed compounds to reverse or slow the progression of tissue calcification in subjects with end-stage renal disease (ESRD):

[0249] This example describes the administration of the disclosed compounds to subjects with ESRD and in stable hemodialysis. Administration of the disclosed compounds will result in changes in aortic compliance (by plethysmography), vascular calcification, and certain biomarker levels that indicate a slowing of the progression of tissue calcification, prevention, or reversal of tissue calcification.

[0250] ESRD subjects undergoing stable hemodialysis receive 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg or 500 mg of the disclosed compound orally once a day for at least 2 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 6 months, 1 year or indefinitely. The dosage form is 5 mg, 10 mg, 25 mg, 50 mg, 75 mg or 100 mg soft gel capsules. Two 50 mg capsules are administered once a day to the 100 mg dose cohort.

[0251] A 50-year-old, 65-kg male patient was diagnosed with typical symptoms associated with moderate calciphylaxis and was treated with 100 mg of compound Ia.1 , m=9, n=7 treatment for 8 weeks. After the treatment period, the patient was admitted and evaluated. The patient was found to have significant changes in the levels of biomarkers examined, indicating a reduction of approximately 10% in vascular calcification and a 10% reduction in tissue calcification.

[0252] A 65-year-old, 45-kg female patient was diagnosed with typical symptoms associated with moderate calciphylaxis and was treated with 20 mg of compound Ia.1 , m=9, n=7 treatment, lasting 10 weeks. After the treatment period, the patient was admitted and evaluated. It was found that the patient's examined biomarker levels had changed significantly, indicating that vascular calcification decreased by about 20%, and tissue calcification also decreased by 15%.

[0253] A 55-year-old, 70-kg male patient was diagnosed with typical symptoms associated with moderate calciphylaxis and was treated with 50 mg of compound Ia.1 , m=9, n=7 treatment, lasting 3 months. After the treatment period, the patient was admitted to the hospital and evaluated. It was found that the patient's examined biomarker levels had changed significantly, indicating that vascular calcification decreased by about 25%, and tissue calcification also decreased by 20%.

[0254] Coronary artery calcium scores (CAC) are used to estimate the extent of calcification in the thoracic arteries. A high CAC score indicates calcification, and the goal of treatment is to prevent a long-term increase in the CAC score, or to reverse its growth, or to slow the rate of increase.

[0255] Aortic plethysmography is also used to measure arterial compliance, which decreases as calcification increases. Pulse wave velocity (PWV) is also measured to assess arterial compliance. The above measurements can be used to assess the effectiveness of treatments designed to prevent, slow the progression of, arrest or reverse vascular calcification. These measurements are used before and after treatment with the disclosed compounds to assess the value of the treatment.

[0256] Several biomarkers can be assessed to determine the efficacy of the disclosed compounds at three dose levels. Exemplary biomarkers include PIVKA-II; uncarboxylated and total matrix Gla protein (MGP); uncarboxylated, carboxylated, and total osteocalcin; uncarboxylated, carboxylated, and total protein C, and hs-CRP. Blood samples are obtained to measure biomarkers, which is most convenient during hemodialysis patient visits.

[0257] Administration of the disclosed compounds may result in (i) a decrease in PIVKA-II, which indicates a delay in the progression of tissue calcification, prevention or reversal of tissue calcification, (ii) a decrease in uncarboxylated MGP, uncarboxylated osteocalcin and / or uncarboxylated protein C, which indicates a delay in the progression of tissue calcification, prevention or reversal of tissue calcification and / or (iii) a decrease in hs-CRP, which indicates a delay in the progression of tissue calcification, prevention or reversal of tissue calcification and / or a reduction in inflammation. Following daily administration of 0.2 mg, 2 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg or 500 mg or more of the disclosed compounds and compositions, at least one of PIVKA-II, incompletely carboxylated matrix Gla protein (MGP), incompletely carboxylated osteocalcin will show changes indicative of a delay in the progression of tissue calcification, prevention or reversal of tissue calcification.

[0258] Thirteen (13) different representative cell lines, using both cell banks and mouse cell models, as well as the HL-60 human leukemia cell line, were used to test the above-mentioned inclusion formula I , II or III The compounds of naphthol derivatives and the biological activities of the compounds disclosed herein are disclosed in the experimental section below. Other cell lines for testing different cancers can be obtained from the American Type Culture Collection (ATCC).

[0259] Study Design:

[0260] Cells were seeded in 96-well microplates and after 3 days of exposure to the compounds disclosed above, the number of viable cells in the culture medium was determined using the CellTiter-Glo Luminescent Cell Viability Kit based on the quantification of the ATP present, which indicates the presence of metabolically active cells. The OD signal is proportional to the amount of ATP present. The amount of ATP is proportional to the number of cells present in the culture medium. This method is used to monitor drug-induced effects on cell proliferation.

[0261] Materials: Reagents and Antibodies:

[0262]

[0263]

[0264] Description of cell lines and media composition:

[0265]

[0266]

[0267] FBS-fetal bovine serum; Dulbecco's modified Eagle's medium (DMEM); Roswell Park Memorial Institute (RPMI) medium 1640.

[0268] SCLC review; see BJ Drapkin et al., Cold Spring Harb. Perspect. Med. 2021; 11:a038240. Compounds were tested at 10 mg in DMSO at a concentration of 100 μM in a 20 mM stock solution using 1 mL of stock solution.

[0269] Preparation of cell assay plates: Day 1

[0270] 1) Pre-warm the culture medium, 0.25% trypsin, and cell culture medium in a 37°C water bath.

[0271] 2) Observe the cells under a microscope to assess confluence and confirm the absence of bacterial and fungal contaminants.

[0272] 3) Remove the culture medium and wash the cells twice with 10 mL of PBS. Add 5 mL of 0.25% trypsin / EDTA reagent to the T-75 flask. Place the flask in an incubator for a few minutes or until the cells detach. Add 10 mL of fresh cell culture medium containing FBS, rinse the cells and transfer to a centrifuge tube. For suspended cells, collect the cells and centrifuge the cells directly.

[0273] 4) The collected cells were centrifuged at 1200 rpm for 5 minutes.

[0274] 5) After centrifugation, discard the supernatant and resuspend the cell pellet with 10 mL of cell culture medium containing FBS.

[0275] 6) Remove 20 μL of the resuspended cells for cell counting. Count the cells using the Cell Counter Star by adding 20 μL of cell suspension to 20 μL of dye, and record the number of live cells and viability in the cell tracking table.

[0276] 7) Use cell culture medium containing FBS and adjust the volume of the suspension to achieve a cell concentration of 3000 cells / well for each adherent cell line. For HL-60, the seeding density is 5000 cells / well.

[0277] 8) According to the plate map, transfer 90 μL of cell suspension to each well of a 96-well white plate. For blank control, add 100 μL of complete medium.

[0278] 9) Incubate the cells overnight at 37°C / 5% CO2.

[0279] Compound dose gradient solution preparation: Day 2

[0280] 1) Dissolve solid compounds in DMSO to form stock solutions.

[0281] 2) Dilute the compound accordingly.

[0282] 3) Add 10 μL of diluted compound at each concentration to 90 μL of cells. For the vehicle group, add 10 μL of the corresponding vehicle to 90 μL of cells.

[0283] 4) For each of the tested compounds and cisplatin, cells were incubated at 37°C / 5% CO2 for 3 days.

[0284] CTG test: Day 5

[0285] 1) Incubate the plate at room temperature and protected from light for 30 minutes.

[0286] 2) Thaw three bottles of CellTiter at room temperature Reagents and equilibrate to room temperature before use. Protect from light.

[0287] 3) Add 100 μL / well of CellTiter to each well. Reagents. Protect from light.

[0288] 4) Mix the contents on an orbital shaker for 2 minutes.

[0289] 5) Incubate the plate at room temperature for 10 minutes to allow the luminescent signal to stabilize.

[0290] 6) Read the plate in Envision.

[0291] Each cell line was plated in 4 plates, for a total of 52 plates.

[0292] The results of the cell assays showed that the disclosed compounds were positive for the changes in viable cells and cell proliferation in the culture medium; and that the activities of some compounds were significantly higher than those of other compounds at different concentrations.

[0293] Topical preparations for wound treatment:

[0294] Mode I , II or III The compounds and compositions containing these compounds can be effectively used as topical compositions or wound dressing compositions for treating wounds or damaged skin, such as wounds associated with tissue calcification and calciphylaxis. The compositions can heal, restore and regenerate wounds and skin of patients. In one variation, the composition comprises about 1.25 mg to 500 mg of formula I , II or III In another variation, the composition may comprise from about 0.01% wt / wt to 20% wt / wt, or about 1%, 5%, 10%, 15%, or about 20% of a compound of formula I , II or III The wound may be an acute wound or a chronic wound. Such wounds may include ulcers such as diabetic ulcers, bedsore ulcers and venous ulcers.

[0295] In one variation, topical application of the composition to damaged skin can reduce inflammation and promote wound healing. Without being bound by any theory claimed herein, it is noteworthy that the composition can stimulate proteasome function and promote normal fibroblast (i.e., reverse aging) activity, as well as the formation of new collagen. This function helps the proteasome to clear fragmented protein particles, including collagen, thereby minimizing the accumulation of undesirable clumping collagen.

[0296] In one variation, the composition can be prepared in the form of a gel, cream or ointment. In another variation, the topical composition further comprises at least one component selected from the group consisting of: water, solvent, preservative, surfactant, gelling agent and pH balancer. The compound or mixture of compounds can be prepared using methods known in the art for use in the form of a pharmaceutical carrier. See, for example, Remington: Pharmaceutical Science and Practice (9th edition, 1995), which is incorporated herein by reference. The compound can be mixed with an acceptable carrier. Available carriers include vaseline, lanolin, polyethylene glycol, alcohol, transdermal enhancers and combinations of two or more thereof. In another variation, the topical composition can be partially or completely incorporated in a delivery vehicle, such as microspheres or nanoparticles, or encapsulated, such as liposomes. In another variation, the topical composition can be pre-impregnated in / on a supporting structure, such as a tape, patch or bandage.

[0297] In another embodiment, the composition is formulated and delivered using a transdermal patch. As provided herein, a transdermal patch provides a formula for a desired period of time. I , II or III The transdermal patch can also be easily removed if the patient experiences undesirable side effects associated with the patch. As formulated in the present invention, the transdermal patch can provide a more precise dose of the compound over time. In one variation, the transdermal patch provides a dose of the formula to a drug delivery area or drug delivery zone (i.e., the area where the composition containing the active agent contacts the intact human skin surface or wound) in the following amount: I , II or III Delivery rate of the compound or mixture thereof: about 1 μg / cm 2 / day to about 600 μg / cm 2 / day, about 1μg / cm 2 / day to about 280 μg / cm 2 / day, about 10μg / cm 2 / day to about 280 μg / cm 2 / day or about 50 μg / cm 2 / day to about 250 μg / cm 2 In one variation, the composition is formulated to deliver the drug continuously over a continuous period of about 5 hours to 7 days. I , II or III In another variation, the transdermal patch is formulated to deliver the compound in a continuous time period of about 12 hours to 24 hours or about 1 day to 3 days. The transdermal patch can be continuously applied in an application time period of about 1 week to about 1 month.

[0298] In addition to the I , II or III In addition to the compound or its mixture, the topical composition can also include other additives, such as pH regulating additives, including acid, such as citric acid or lactic acid, alkali or buffer, such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate or sodium gluconate. In addition, in some embodiments, the composition can contain an antimicrobial preservative. The antimicrobial preservative includes, for example, methylparaben, propylparaben, benzyl alcohol, ethylhexylglycerol, potassium sorbate, phenoxyethanol, EDTA, grapefruit seed extract, tea tree oil, sodium benzoate, dehydroacetic acid and a combination thereof. In another variation, the composition can include an antifungal preservative, optionally in combination with an antibacterial preservative. In a variation, the composition can include an anti-biofilm antimicrobial, such as lactoferrin, xylitol, gallium farnesol, dispersant B and EDTA or a combination thereof.

[0299] In one embodiment, the skin patch can be prepared as follows: 1) a metered amount of formula I , II or III 1) The compound or mixture of compounds of formula (I) is dissolved in a solvent such as THF, EtOAc or methanol or a mixture thereof and added to the DuroTak 87-2194 adhesive to form a solution containing the compound. 2) The solution of the compound and adhesive blend is formed on a release liner using a mechanical coater. 3) The coated release liner is then passed through an oven to evaporate the solvent (e.g., ethyl acetate and any solvent present in the liquid DuroTak) to form a solid viscous adhesive matrix layer containing the formula (I) dispersed in the adhesive matrix. I , II or III 4) The polyethylene film is then laminated to the adhesive matrix. 5) The compound-containing patch laminate is then cut into specified sizes using a die cutter, and the patches are then individually packaged in a sealed foil liner material.

[0300] In another variation, the composition can be used without a microporous tape or other support structure. In one variation, the composition is sufficiently viscous or sticky to create an occlusive dressing without a support structure such as tape. In another variation, the composition can be applied under a support structure or applied on top of a support structure.

[0301] In another variation, if the composition is formulated in a microporous support structure, such as a tape, for receiving the composition on an outer surface of the tape and contacting the damaged skin through the pores, the skin or damaged skin tissue delivers the composition.

[0302] In one embodiment, tape (or other support structure) is applied to the length of the wound or scar, and the topical composition is applied to the tape. After the wound is observed or before the subcutaneous wound breaks the skin, the use of the tape and gel can be started as soon as possible. In a variation, the tape is a microporous tape that allows the topical composition to contact the skin when the topical composition is applied on the tape. In a variation, the topical composition can be applied to the skin before the tape is placed on the composition.

[0303] Therefore, the present application discloses a method for accelerating wound healing, the method comprising applying a therapeutically effective amount of a formula I , II or III Compounds to promote wound healing. Topical administration can be with the formula I , II or III The oral administration of the composition is combined to accelerate wound healing, which means that the topical application can be combined with the oral administration. I , II or III The invention can be administered before, during and / or after administration of a compound of the invention, as disclosed herein.

[0304] As described herein, acceleration of wound healing includes increasing the rate of healing of wounds with or without oral administration compared to the rate of healing of untreated wounds. I , II or III The accelerated wound healing achieved using the disclosed methods can result in a treated wound that heals at least 5% faster, at least 10%, 15%, 20%, 25% faster; or at least 50%, 75% or 100% faster than an untreated wound. This at least 5% to 100% increase or acceleration in wound healing can be combined with oral administration of a wound healing agent. I , II or III The compounds are compared to patients or subjects being treated concurrently.

[0305] Although a number of exemplary embodiments, aspects and variations have been provided herein, those skilled in the art will recognize certain modifications, permutations, additions and combinations of the embodiments, aspects and variations, as well as certain sub-combinations of the embodiments, aspects and variations. It is intended that the following claims be interpreted as including within their scope all such modifications, permutations, additions and combinations of the embodiments, aspects and variations, as well as certain sub-combinations of the embodiments, aspects and variations. The entire disclosures of all documents cited in this application are incorporated herein by reference.

[0306] References: 1) Rachel M. Holden et al., Vitamins K and D Status in Stages 3-5 Chronic Kidney Disease; Clin J Am Soc Nephrol 5:590-597, 2010. 2) Pilkey, RMMD et al., Subclinical Vitamin K Deficiency in Hemodialysis Patients; Am J Nephrol 49:432-439, 2007. 3) Westhofen P et al., Human vitamin K 2,3-epoxide reductase complex subunit 1-like 1 (VKORC1L1) mediates vitamin K-dependent intracellular antioxidant function. function). J Biol Chem 2011; 286: 15085-94. 4) Caspers, M. et al., Two enzymes catalyze vitamin K 2,3-epoxide reductase activity in mouse: VKORC1 is highly expressed in exocrine tissues while VKORC1L1 is highly expressed in brain. Thrombosis Research 135: 977-983, 2015. 5) Himmelfarb, J. et al., Plasma protein thiol oxidation and carbonylformation in chronic renal failure. Kidney International, Vol. 58: 2571-2578 2000.6) Price, P. A. et al., Discovery of a High Molecular Weight Complex of Calcium, Phosphate, Fetuin, and Matrix-Carboxyglutamic Acid Protein in the Serum of Etidronate-treated Rats. Journal of Biological Chemistry 277(6):3926-3934, 2002. 7) Pasch, A. et al., Nanoparticle-Based Test Measures Overall Propensity for Calcification in Serum. Journal of the American Society of Nephrology 23:1744-1752, 2012. 8) Nigwekar, S. U. et al., Vitamin K-Dependent Carboxylation of Matrix Gla Protein Influences the Risk of Calcification Defense. Calciphylaxis). Journal of the American Society of Nephrology 28:1717-1722, 2017.

Claims

1. A biologically active naphthol derivative having the formula I , II or III : in: m is an integer from 2 to 15; m 1 an integer from 3 to 15 when present; n is 7, 8, 9, or 10; R is H, or is selected from the group consisting of: -C(O)C1-C6 alkyl, -C(O)(CH2) q -OC(O)C1-C6 alkyl and -Z-(C=Y)(CH2) s -(OCH2CH2)-OR 1 ; R 1 is H or -CH3; R 2 is H or C1-C3 alkyl; q is 1, 2, or 3; s is 1, 2, or 3; Y is O, NR 2 or S; Z is -C(O)CH2O-, -C(O)CH2NH- or absent; and Z 1 It is -C(O)CH2O-, -C(O)CH2NH- or does not exist.

2. The naphthol derivative according to claim 1, which has the formula Ia , Ib or Ic : in: m is an integer from 2 to 15; n is 7, 8, 9, or 10; R is H, or is selected from the group consisting of: -C(O)C1-C6 alkyl, -C(O)(CH2) q -OC(O)C1-C6 alkyl and -Z-(C=Y)(CH2) s -(OCH2CH2)-OR 1 ; R 1 is H or -CH3; R 2 is H or C1-C3 alkyl; q is 1, 2, or 3; s is 1, 2, or 3; Y is O, NR 2 or S; and Z is -C(O)CH2O-, -C(O)CH2NH- or absent.

3. The naphthol derivative according to claim 1, which has the formula Ia.1 , Ib.1 or Ic.1 : in: m is an integer from 8 to 12; n is 7, 8, 9 or 10; and R 1 It is H or -CH3.

4. The naphthol derivative according to claim 3, which has the formula IIa , IIb or IIc : in: m is an integer from 8 to 12; n is 7, 8, 9, or 10; s is 1, 2, or 3; and R 1 It is H or -CH3.

5. The naphthol derivative according to claim 1, which has the formula Ia.1 , Ib.1 , Ic.1 , Ia.2 , Ib.2 , Ic.2 , IIa , IIb , IIc , IIa.1 , IIb.1 , IIc.1 , IIa.1.1 , IIb.1.1 , IIc.1.1 , IIa.1.2 , IIb.1.2 , IIc.1.2 , IIa.2 , IIb.2 , IIc.2 , IIa.3 , IIb.3 , IIc.3 , IIa.4 , IIb.4 , IIc.4 , IIa.5 , IIb.5 and IIc.5 : 6 . The naphthol derivative according to claim 1 , wherein n is 7.

7. A pharmaceutical composition comprising a therapeutically effective amount of the naphthol derivative or a mixture thereof according to any one of claims 1 to 6; and a pharmaceutically acceptable excipient, wherein the composition is effective in treating a condition associated with vitamin K selected from osteoporosis, atherosclerosis, calciphylaxis or tissue calcification.

8. A method for increasing the tissue concentration of naphthol as a cofactor for gamma-glutamate carboxylase (GGCX), which is used to catalyze the carboxylation of vitamin K-dependent proteins associated with the treatment or prevention of osteoporosis, atherosclerosis, calciphylaxis or tissue calcification in patients in need thereof, the method comprising administering a therapeutically effective amount of a naphthol derivative, or a pharmaceutical composition comprising an effective amount of a naphthol derivative according to any one of claims 1 to 7 or a mixture thereof.

9. A method for treating a disease in a mammal, the disease being selected from the group consisting of: neurodegenerative diseases, proliferative diseases, retinopathy, rheumatoid polyarthritis, atherosclerosis, amyotrophic lateral sclerosis, cerebral ischemia, cataracts, systemic infections, pathologies associated with skin aging and tissue aging, pathologies associated with mitochondrial dysfunction, cachexia associated with malnutrition, wherein the treatment is associated with an extension of the life span of the mammal, the method comprising administering a therapeutically effective amount of a compound or composition comprising a naphthol derivative or a mixture thereof according to any one of claims 1 to 7.

10. A method for treating a mammal suffering from a disease selected from the group consisting of vitamin K deficiency, osteoporosis, proliferative diseases and cardiovascular diseases, the method comprising administering to the mammal a therapeutically effective amount of a naphthol derivative according to any one of claims 1 to 7 or a mixture thereof.

11. The method of claim 9, wherein the proliferative disease is selected from the group consisting of cancer, leukemia, and inflammatory disease.

12. A method for treating or preventing osteoporosis and / or osteopenia, the method comprising administering to a patient in need of treatment a therapeutically effective amount of a composition comprising a naphthol derivative or a mixture thereof according to any one of claims 1 to 7.

13. A method for treating, preventing, delaying the progression of, arresting and / or reversing calciphylaxis in a mammal in need thereof, the method comprising administering to the mammal a therapeutically effective amount of a composition comprising a substantially pure naphthol derivative or a mixture thereof according to any one of claims 1 to 7 and a pharmaceutically acceptable excipient to prevent, delay the progression of, arrest or reverse calciphylaxis.

14. The method of claim 13, wherein the mammal suffers from distal calciphylaxis and / or central calciphylaxis.

15. The method of claim 13 or 14, wherein the mammal suffers from diabetes, chronic kidney disease or end-stage renal disease.

16. The method of claim 15, wherein the mammal has stage 3, stage 4, or stage 5 chronic kidney disease.

17. The method of any one of claims 9 to 16, wherein the mammal is undergoing hemodialysis.

18. The method of any one of claims 9 to 17, wherein the mammal is receiving non-warfarin based anticoagulant therapy.

19. The method of claim 18, wherein the anticoagulant therapy is oral anticoagulant therapy.

20. The method of claim 18, wherein the anticoagulant therapy comprises an inhibitor of factor Xa activity selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban, or fondaparinux; or an inhibitor of factor IIa activity selected from dabigatran or argatroban.

21. The method of any one of claims 8 to 20, wherein the mammal suffers from chronic obstructive pulmonary disease (COPD).

22. The method of any one of claims 8 to 21, wherein the mammal suffers from a calciphylaxis-related skin lesion.

23. The method of claim 22, wherein administering the composition reduces the total surface area of ​​the skin lesions by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

24. The method of any one of claims 8 to 23, wherein administration of a compound according to any one of claims 1 to 6, or a mixture thereof, to the mammal increases the serum T50 value of the mammal by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% relative to the serum T50 value of the mammal prior to administration of a compound according to any one of claims 1 to 6, or a mixture thereof.

25. The method of any one of claims 13 to 24, wherein administration of a compound according to any one of claims 1 to 6 or a mixture thereof increases the ratio of carboxylated vitamin K-dependent proteins to non-carboxylated vitamin K-dependent proteins in the plasma of the mammal, the ratio after administration of the composition being greater than the ratio before administration of the composition.

26. A method for treating, preventing, slowing the progression of, preventing and / or reversing tissue calcification in a prediabetic mammal (or subject) having diabetes, chronic kidney disease or a combination thereof and in need thereof, the method comprising administering to the mammal at least 50 mg per day of a compound or mixture thereof according to any one of claims 1 to 6 to prevent, slow the progression of and / or prevent tissue calcification, wherein the compound or mixture thereof according to any one of claims 1 to 6 is administered in the form of a pharmaceutical composition.

27. The method of claim 26, wherein the mammal suffers from diabetes.

28. The method of claim 27, wherein the mammal suffers from type II diabetes.

29. The method of claim 26, wherein the mammal has been diagnosed with prediabetes.

30. The method of any one of claims 26 to 29, wherein the mammal suffers from chronic kidney disease.

31. The method of any one of claims 26 to 30, wherein the mammal has stage 4 or stage 5 chronic kidney disease / end stage renal disease.

32. The method of any one of claims 26 to 31, wherein the mammal is undergoing hemodialysis.

33. The method of any one of claims 26 to 32, wherein the mammal is receiving non-warfarin based anticoagulant therapy.

34. The method of claim 33, wherein the anticoagulant therapy is oral anticoagulant therapy.

35. The method of claim 33, wherein the anticoagulant therapy comprises an inhibitor of factor Xa activity selected from apixaban, rivaroxaban, betrixaban, edoxaban, omixaban, ritoxaban, irrigaban, or fondaparinux; or an inhibitor of factor IIa activity selected from dabigatran or argatroban.

36. A method for treating, preventing, slowing the progression of, arresting and / or reversing tissue calcification in a mammal undergoing hemodialysis and in need thereof, the method comprising administering to the mammal at least 5 mg of a compound according to any one of claims 1 to 6 or a mixture thereof per day, thereby preventing, slowing the progression of, arresting and / or reversing tissue calcification, wherein the compound according to any one of claims 1 to 6 or a mixture thereof is administered in the form of a pharmaceutical composition.

37. The method of claim 36, wherein the mammal suffers from diabetes.

38. A method for increasing at least one of the bioavailability and serum half-life of: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9, or menaquinone-10 / naphthol-10, the method comprising administering or supplementing a mammal in need thereof with a compound or composition thereof according to any one of claims 1 to 6, wherein the method increases the bioavailability and serum half-life of: menaquinone-7 / naphthol-7, menaquinone-8 / naphthol-8, menaquinone-9 / naphthol-9, or menaquinone-10 / naphthol-10 by 5% when compared to administration or supplementation with menaquinone-7, menaquinone-8, menaquinone-9, or menaquinone-10.

39. A method for treating at least one of a wound or a pain and inflammation area caused by calciphylaxis or tissue calcification in a subject in need thereof, the method comprising topically applying a dermal structure or a transdermal patch to the wound or pain and inflammation area of ​​the subject, wherein the dermal structure or the transdermal patch comprises a compound according to any one of claims 1 to 6 to treat the wound, pain and inflammation area.

40. The method of claim 39, wherein the topical application of the dermal structure or the transdermal patch is performed before, in conjunction with (or simultaneously with) or after the method of any one of claims 8 to 38.

41. The method of claim 11, wherein the cancer is selected from the group consisting of: Melanoma, lung cancer, breast cancer, leukemia, neuroblastoma, glioblastoma, cervical cancer, liver cancer (hepatocellular carcinoma, HCC), colorectal cancer, pancreatic cancer, bladder cancer, kidney cancer, prostate cancer, ovarian cancer and head and neck cancer.

42. A method for treating, preventing, delaying the progression of, arresting and / or reversing Alzheimer's disease (AD) in a mammal or subject in need thereof, the method comprising administering to the mammal or subject at least 5 mg of a compound according to any one of claims 1 to 6 or a mixture thereof per day, thereby preventing, delaying the progression of, arresting and / or reversing Alzheimer's disease, wherein the compound according to any one of claims 1 to 6 or a mixture thereof is administered in the form of a pharmaceutical composition.

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